Fatty acid formulations and methods of use thereof

ABSTRACT

The present invention provides dietary formulations comprising polyunsaturated fatty acids and vitamin E. The present invention further provides methods of treating various conditions, generally involving administering to an individual in need thereof an effective amount of a subject dietary formulation.

CROSS-REFERENCE

This application is a divisional of U.S. patent application Ser. No.15/424,358, now U.S. Pat. No. 10,398,671, which is a continuation ofU.S. patent application Ser. No. 13/439,192, filed Apr. 4, 2012, nowU.S. Pat. No. 9,687,016, which is a divisional of U.S. patentapplication Ser. No. 12/033,431, filed Feb. 19, 2008, now U.S. Pat. No.10,300,034, which claims the benefit of U.S. Provisional PatentApplication No. 60/891,097, filed Feb. 22, 2007, and of U.S. ProvisionalPatent Application No. 60/981,262, filed Oct. 19, 2007, whichapplications are incorporated herein by reference in their entirety.

BACKGROUND

Apraxia of speech is a neurologically based motor planning disorder ofunknown etiology. Approximately half of children with autistic spectrumdisorders have some degree of apraxia, although not all apraxic childrenare autistic. Children with apraxia find it very difficult to correctlypronounce sounds, syllables, and words. Some children remain speechlessand require the use of augmentative communication devices or utilize apicture exchange communication system. Many children with apraxiapresent with homogeneous symptoms of neurological dysfunction thataffect coordination, muscle tone, and sensory issues in addition tospeech production difficulties, suggesting a common underlying mechanismof disease in these children. Standard treatment generally involvesspeech therapy with a speech pathologist knowledgeable in apraxia.

There is a need in the art for alternative approaches to treatment ofapraxia.

LITERATURE

U.S. Pat. Nos. 5,776,978; 6,914,073; 7,169,385; 6,184,251.

SUMMARY OF THE INVENTION

The present invention provides dietary formulations comprisingpolyunsaturated fatty acids and vitamin E. The present invention furtherprovides methods of treating various conditions, generally involvingadministering to an individual in need thereof an effective amount of asubject dietary formulation.

Definitions

“Fatty acids” refer to a family of carboxylic acids having a hydrocarbonchain of from about 12 to about 24 carbons in length. Unsaturated fattyacids have at least one carbon-carbon double bond in the hydrocarbonchain. Unsaturated fatty acids include monounsaturated fatty acids andpolyunsaturated fatty acids (PUFAs). Unsaturated fatty acids aredesignated by the position of the first double bond from the methyl endof the hydrocarbon chain. Omega-3 fatty acids have a first double bondat the third carbon from the methyl end of the chain; and include, e.g.,α-linolenic acid (octadeca-9,12,15-trienoic acid), stearidonic acid(octadeca-6,9,12,15-tetraenoic acid), eicosapentaenoic acid(eicosa-5,8,11,14,17-pentaenoic acid; “EPA”), docosapentaenoic acid(docosa-7,10,13,16,19-pentaenoic acid), eicosatetraenoic acid(eicosa-8,11,14,17-tetraenoic acid), and docosahexaenoic acid(docosa-4,7,10,13,16,19-hexaenoic acid; “DHA”). Omega-6 fatty acids havea first double bond at the sixth carbon from the methyl end of thechain; and include, e.g., linoleic acid (9,12-octadecadienoic acid),γ-linolenic acid (6,9,12-octadecatrienoic acid; GLA), eicosadienoic acid(11,14-eicosadienoic acid), dihomo-γ-linolenic acid(8,11,14-eicosatrienoic acid), arachidonic acid(5,8,11,14-eicosatetraenoic acid), docosadienoic acid(13,16-docosadienoic acid), adrenic acid (7,10,13,16-docosatetraenoicacid), docosapentaenoic acid (4,7,10,13,16-docosapentaenoic acid), andcalendic acid (8E,10E,12Z-octadecatrienoic acid), and the like. Omega-9fatty acids have a first double bond at the ninth carbon from the methylend of the chain; and include, e.g., oleic acid (cis-9-octadecenoicacid); eicosenoic acid (cis-11-eicosenoic acid); mead acid(all-cis-5,8,11-eicosatrienoic acid); erucic acid (cis-13-docosenoicacid); and nervonic acid (cis-15-tetracosenoic acid).

As used herein, “vitamin E” refers to a family of eight molecules havinga chromanol ring (chroman ring with an alcoholic hydroxyl group) and a12-carbon aliphatic side chain containing two methyl groups in themiddle and two more methyl groups at the end. The side chain of thetocopherols is saturated, while the side chain of the tocotrienolscontain three double-bonds, all of which adjoin a methyl group. Thetocopherols and the tocotrienols exist in four isoforms, referred to asalpha, beta, gamma and delta isoforms. The isoforms are named on thebasis of the number and position of the methyl groups on the chromanolring. The alpha form has three methyl groups, the beta and gamma formshave two methyl groups and the delta for has only one methyl group. Asused herein, “vitamin E” refers to one or more of α-tocopherol,β-tocopherol, γ-tocopherol, α-tocotrienol, β-tocotrienol, andγ-tocotrienol. “Vitamin E” also includes esters of a vitamin E isoform.For example, “vitamin E” includes esters of a tocopherol, includingacetates and succinates.

As used herein, the term “lipoic acid” refers to α-lipoic acid, which isa chiral molecule also known as thioctic acid; 1,2-diethylene-3pentanoic acid; 1,2-diethylene-3 valeric acid; and 6,8-thioctic acid.Unless specified the term “lipoic acid” encompasses the racemic mixtureas well as any other (non-50/50) mixture of the enantiomers includingsubstantially pure forms of either the R-(+) or the S-(−) enantiomer.Further, unless specified otherwise the term covers pharmaceuticallyacceptable salts (e.g. Na and K salts) and amides, esters andmetabolites of the acid. The molecule formula is C₈H₁₄O₂S₂ the molecularweight is 206.32 and it has a pKa of 4.7. In referring topharmaceutically acceptable salts, the term is intended to encompass aconventional term of pharmaceutically acceptable acid addition saltswhich refer to salts which retain the biological effectiveness andproperties of the free-base form of the acid and which are notbiologically or otherwise undesirable, formed with inorganic acids suchas hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like, and organic acids such as acetic acid,propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid,malconic acid, succinic acid, maleic acid, fumaric, tartaric acid,citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonicacid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid andthe like. The same is true with respect to amides, esters andmetabolites which can be formed and maintain biological effectivenessand not have significant undesirable biological properties.

“Carnitine” is also known as3-carboxy-2-hydroxy-N,N,N-trimethyl-1-propanaminium hydroxide, innersalt; (3-carboxy-2-hydroxypropyl)trimethylammonium hydroxide, innersalt; gamma-amino-beta-hydroxybutyric acid trimethylbetaine;gamma-trimethyl-beta-hydroxybutyrobetaine;3-hydroxy-4-(trimethyl-ammonio)butanoate. As used herein, the term“carnitine” includes carnitine and “carnitine analogs” and encompassesracemic or essentially pure L-carnitine (carnitine), or a correspondingalkanoyl-carnitine such as acetyl-carnitine or propionyl-carnitine, or asuitable salt of such compounds such as L-carnitine tartrate,L-carnitine fumarate, L-carnitine-magnesium-citrate, acetyl-L-carnitinetartrate, acetyl-L-carnitine-magnesium-citrate, or any mixture of theaforementioned compounds.

As used herein, “pharmaceutically acceptable carrier” includes anymaterial which, when combined with an active ingredient of acomposition, allows the ingredient to retain biological activity andwithout causing disruptive reactions with the subject's immune system.Examples include, but are not limited to, any of the standardpharmaceutical carriers such as a phosphate buffered saline solution,water, emulsions such as oil/water emulsion, and various types ofwetting agents. Exemplary diluents for aerosol or parenteraladministration are phosphate buffered saline or normal (0.9%) saline.Compositions comprising such carriers are formulated by well knownconventional methods (see, for example, Remington's PharmaceuticalSciences, Chapter 43, 14th Ed. or latest edition, Mack Publishing Co.,Easton Pa. 18042, USA; A. Gennaro (2000) “Remington: The Science andPractice of Pharmacy”, 20th edition, Lippincott, Williams, & Wilkins;Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) H. C. Anselet al., eds 7^(th) ed., Lippincott, Williams, & Wilkins; and Handbook ofPharmaceutical Excipients (2000) A. H. Kibbe et al., eds., 3^(rd) ed.Amer. Pharmaceutical Assoc.

As used herein, the terms “treatment,” “treating,” and the like, referto obtaining a desired pharmacologic and/or physiologic effect. Theeffect may be prophylactic in terms of completely or partiallypreventing a disease or symptom thereof and/or may be therapeutic interms of a partial or complete cure for a disease and/or adverse affectattributable to the disease. “Treatment,” as used herein, covers anytreatment of a disease in a mammal, particularly in a human, andincludes: (a) reducing the incidence and/or risk of relapse of thedisease during a symptom-free period; (b) relieving or reducing asymptom of the disease; (c) preventing the disease from occurring in asubject which may be predisposed to the disease but has not yet beendiagnosed as having it; (d) inhibiting the disease, i.e., arresting itsdevelopment (e.g., reducing the rate of disease progression); (e)reducing the frequency of episodes of the disease; and (f) relieving thedisease, i.e., causing regression of the disease.

The terms “individual,” “host,” “subject,” and “patient,” usedinterchangeably herein, refer to a mammal, e.g., a human. Where the hostis a mammal, the subject will generally be a human, but may also be adomestic livestock (e.g., horse, cattle, pigs, goats, sheep, etc.), amammalian laboratory subject (e.g., a rodent, a lagomorph, etc.), ormammalian pet animal.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. Thus, for example, reference to “anomega-3 fatty acid” includes a plurality of such fatty acids andreference to “the vitamin E isoform” includes reference to one or morevitamin E isoforms and equivalents thereof known to those skilled in theart, and so forth. It is further noted that the claims may be drafted toexclude any optional element. As such, this statement is intended toserve as antecedent basis for use of such exclusive terminology as“solely,” “only” and the like in connection with the recitation of claimelements, or use of a “negative” limitation.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

DETAILED DESCRIPTION

The present invention provides dietary formulations comprisingpolyunsaturated fatty acids (PUFA) and vitamin E. A subject formulationis useful for treating various motor and communication disorders as wellas allergic disorders. The present invention thus provides methods oftreating such disorders, generally involving administering to anindividual in need thereof an effective amount of a subject dietaryformulation.

The present dietary formulations include a PUFA (e.g., an omega-3 fattyacid) and vitamin E in amounts and proportions that provide fortreatment of the various categories of disorders mentioned above,including, e.g., apraxia. It has been found that such dietaryformulations provide for a reduction in symptoms of various motor andcommunication disorders. Insights into a possible common mechanismcontributed to the development of the subject formulations. While notwishing to be bound by theory, it is possible that dysregulation offatty acid metabolism may play a central role in disorders such asapraxia, autism, and other disorders characterized at least in part bymotor and/or communication dysfunction. In addition, it is possible thatabnormal vitamin E bioavailability may play a role in these disorders.Furthermore, abnormal carnitine bioavailability may contribute todysregulation of fatty acid metabolism on the cellular and mitochondriallevel. Inflammation, oxidative stress and depletion of antioxidants suchas glutathione in addition to vitamin E (vit E) may contribute to thesedisorders.

Formulations

As noted above, a subject dietary formulation comprises at least onepolyunsaturated fatty acid (PUFA) and at least one vitamin E isoform.Suitable PUFA include, but are not limited to, omega-3 fatty acids andomega-6 fatty acids. Suitable omega-3 fatty acids include, e.g.,α-linolenic acid (octadeca-9,12,15-trienoic acid), stearidonic acid(octadeca-6,9,12,15-tetraenoic acid), eicosapentaenoic acid(eicosa-5,8,11,14,17-pentaenoic acid; “EPA”), docosapentaenoic acid(docosa-7,10,13,16,19-pentaenoic acid), eicosatetraenoic acid(eicosa-8,11,14,17-tetraenoic acid), and docosahexaenoic acid(docosa-4,7,10,13,16,19-hexaenoic acid; “DHA”). Suitable omega-6 fattyacids include, e.g., linoleic acid (9,12-octadecadienoic acid),γ-linolenic acid (6,9,12-octadecatrienoic acid; GLA), eicosadienoic acid(11,14-eicosadienoic acid), dihomo-γ-linolenic acid(8,11,14-eicosatrienoic acid), arachidonic acid(5,8,11,14-eicosatetraenoic acid), docosadienoic acid(13,16-docosadienoic acid), adrenic acid (7,10,13,16-docosatetraenoicacid), docosapentaenoic acid (4,7,10,13,16-docosapentaenoic acid), andcalendic acid (8E,10E,12Z-octadecatrienoic acid).

A subject formulation can comprise one, two, three, four, five, six,seven, or eight different vitamin E isoforms. For example, in someembodiments, a subject formulation comprises α-tocopherol andγ-tocopherol; and substantially no other vitamin E isoforms. In otherembodiments, a subject formulation includes α-tocopherol, γ-tocopherol,and at least one other vitamin E isoform. For example, in someembodiments, a subject formulation includes α-tocopherol, γ-tocopherol,and at least one of β-tocopherol, δ-tocopherol, α-tocotrienol,β-tocotrienol, γ-tocotrienol, and δ-tocotrienol. In other embodiments, asubject formulation includes α-tocopherol, γ-tocopherol, β-tocopherol,δ-tocopherol, α-tocotrienol, β-tocotrienol, γ-tocotrienol, andδ-tocotrienol.

In some embodiments, a subject formulation comprises at least oneomega-3 fatty acid; and at least one vitamin E isoform. In someembodiments, a subject formulation comprises two different omega-3 fattyacids. In some embodiments, a subject formulation compriseseicosapentaenoic acid (EPA); docosohexaenoic acid (DHA); and at leastone vitamin E isoform. In these embodiments, the ratio of EPA to DHA canrange from about 1.5:1 (EPA:DHA) to about 5:1 (EPA:DHA), where the ratiois on a weight basis. For example, the EPA:DHA ratio can range fromabout 1.5:1 to about 2:1, from about 2:1 to about 2.5:1, from about2.5:1 to about 3:1, from about 3:1 to about 3.5:1, from about 3.5:1 toabout 4:1, from about 4:1 to about 4.5:1, or from about 4.5:1 to about5:1. In some embodiments, the EPA:DHA ratio is 2.5:1.

In some embodiments, a subject formulation comprises at least twodifferent omega-3 fatty acids; at least one vitamin E isoform; and atleast one omega-6 fatty acid. For example, in some embodiments, asubject formulation comprises EPA; DHA; and at least one omega-6 fattyacid. In some embodiments, a subject formulation comprises EPA; DHA; atleast one vitamin E isoform; and γ-linolenic acid (GLA). In someembodiments, a subject formulation comprises EPA; DHA; α-tocopherol;γ-tocopherol; and GLA.

In some embodiments, a subject formulation comprises at least twodifferent omega-3 fatty acids; at least one vitamin E isoform; and anomega-9 fatty acid. In some embodiments, a subject formulation comprisesat least two different omega-3 fatty acids; at least one vitamin Eisoform; an omega-6 fatty acid; and an omega-9 fatty acid. The omega-9fatty acid is in some embodiments a monounsaturated fatty acid. In otherembodiments, the omega-9 fatty acid is a PUFA. In some embodiments, theomega-9 fatty acid is oleic acid.

Lipoic Acid

In some embodiments, a subject formulation further comprises α-lipoicacid. Thus, e.g., in some embodiments, a subject formulation comprises aPUFA; at least one vitamin E isoform; and α-lipoic acid. In someembodiments, a subject formulation comprises at least one omega-3 fattyacid; at least one vitamin E isoform; and α-lipoic acid. For example, insome embodiments, a subject formulation comprises EPA; DHA; at least onevitamin E isoform; and α-lipoic acid. In some embodiments, a subjectformulation comprises EPA; DHA; α-tocopherol; γ-tocopherol; and α-lipoicacid. In other embodiments, a subject formulation comprises at least oneomega-3 fatty acid; an omega-6 fatty acid; at least one vitamin Eisoform; and α-lipoic acid. For example, in some embodiments, a subjectformulation comprises EPA; DHA; GLA; α-tocopherol; γ-tocopherol; andα-lipoic acid.

The α-lipoic can exist as a 50/50 or racemic mixture of R-(+)-α-lipoicacid and S-(−)-α-lipoic acid. The α-lipoic acid ingredient of a subjectformulation can be 100% R-(+) enantiomer. However, the α-lipoic acid canbe present in a subject formulation in any mixture of the twoenantiomers e.g. 10% S-(−) and 90% R-(+); 25% S-(−) and 75% R-(+); etc.

Carnitine

In some embodiments, a subject formulation further comprises carnitine.Thus, e.g., in some embodiments, a subject formulation comprises a PUFA;at least one vitamin E isoform; and carnitine. In some embodiments, asubject formulation comprises at least one omega-3 fatty acid; at leastone vitamin E isoform; and carnitine. For example, in some embodiments,a subject formulation comprises EPA; DHA; at least one vitamin Eisoform; and carnitine. In some embodiments, a subject formulationcomprises EPA; DHA; α-tocopherol; γ-tocopherol; and carnitine. In otherembodiments, a subject formulation comprises at least one omega-3 fattyacid; an omega-6 fatty acid; at least one vitamin E isoform; andcarnitine. For example, in some embodiments, a subject formulationcomprises EPA; DHA; GLA; α-tocopherol; γ-tocopherol; and carnitine. Inother embodiments, a subject formulation comprises at least one omega-3fatty acid; an omega-6 fatty acid; an omega-9 fatty acid; at least onevitamin E isoform; and carnitine. For example, in some embodiments, asubject formulation comprises EPA; DHA; GLA; oleic acid; α-tocopherol;γ-tocopherol; and carnitine.

In some embodiments, the carnitine component of a subject formulation is90%-100% L-carnitine (or a salt thereof). In other embodiments, thecarnitine component of a subject formulation is 90%-100%acetyl-carnitine. In other embodiments, the carnitine component of asubject formulation is a mixture of L-carnitine (or a salt thereof) andacetyl-carnitine. For example, in some embodiments, the carnitinecomponent of a subject formulation can comprise 10% L-carnitine and 90%acetyl-carnitine; 15% L-carnitine and 85% acetyl-carnitine; 20%L-carnitine and 80% acetyl-carnitine; 25% L-carnitine and 75%acetyl-carnitine; 30% L-carnitine and 70% acetyl-carnitine; 40%L-carnitine and 60% acetyl-carnitine; 50% L-carnitine and 50%acetyl-carnitine; 60% L-carnitine and 40% acetyl-carnitine; 70%L-carnitine and 30% acetyl-carnitine; 75% L-carnitine and 25%acetyl-carnitine; 80% L-carnitine and 20% acetyl-carnitine; 85%L-carnitine and 15% acetyl-carnitine; or 90% L-carnitine and 10%acetyl-carnitine; or L-carnitine and acetyl-carnitine in any otherproportion, where the percentages are by weight.

Amounts

The amounts in a subject formulation of PUFA and vitamin E, as well asthe amounts of additional components such as carnitine and α-lipoicacid, can vary according to various factors, including, e.g., the age ofthe individual, the weight of the individual, the genetic makeup of theindividual, and the severity of symptoms exhibited by the individual towhom a subject formulation is administered. The following are generalguidelines. Amounts are given as per unit dose.

Where a subject formulation includes omega-3 fatty acids, the omega-3fatty acids are present in amount of from about 100 mg to about 5000 mg,e.g., from about 100 mg to about 200 mg, from about 200 mg to about 300mg, from about 300 mg to about 400 mg, from about 400 mg to about 500mg, from about 500 mg to about 600 mg, from about 600 mg to about 700mg, from about 700 mg to about 800 mg, from about 800 mg to about 900mg, from about 900 mg to about 1000 mg, from about 1000 mg to about 1100mg, from about 1100 mg to about 1200 mg, from about 1200 mg to about1300 mg, from about 1300 mg to about 1400 mg, from about 1400 mg toabout 1500 mg, from about 1500 mg to about 1600 mg, from about 1600 mgto about 1700 mg, from about 1700 mg to about 1800 mg, from about 1800mg to about 1900 mg, from about 1900 mg to about 2000 mg, from about2000 mg to about 2500 mg, from about 2500 mg to about 3000 mg, fromabout 3000 mg to about 3500 mg, from about 3500 mg to about 4000 mg,from about 4000 mg to about 4500 mg, or from about 4500 mg to about 5000mg per unit dose, where the amounts given are for individual omega-3fatty acids or for total omega-3 fatty acids (e.g., where more than oneomega-3 fatty acid is present).

For example, in some embodiments, a subject formulation comprises theomega-3 fatty acids EPA and DHA. In some embodiments, a subjectformulation will comprise EPA in an amount of from about 500 mg to about3000 mg, e.g., from about 500 mg to about 600 mg, from about 600 mg toabout 700 mg, from about 700 mg to about 800 mg, from about 800 mg toabout 900 mg, from about 900 mg to about 1000 mg, from about 1000 mg toabout 1500 mg, from about 1500 mg to about 2000 mg, from about 2000 mgto about 2500 mg, or from about 2500 mg to about 3000 mg per unit dose;and will comprise DHA in an amount of from about 100 mg to about 400 mg,e.g., from about 100 mg to about 150 mg, from about 150 mg to about 200mg, from about 200 mg to about 250 mg, from about 250 mg to about 300mg, from about 300 mg to about 350 mg, or from about 350 mg to about 400mg per unit dose.

Where a subject formulation comprises the omega-3 fatty acids EPA andDHA, the ratio of EPA to DHA can range from about 1.5:1 (EPA:DHA) toabout 5:1 (EPA:DHA), where the ratio is on a weight basis. For example,the EPA:DHA ratio can range from about 1.5:1 to about 2:1, from about2:1 to about 2.5:1, from about 2.5:1 to about 3:1, from about 3:1 toabout 3.5:1, from about 3.5:1 to about 4:1, from about 4:1 to about4.5:1, or from about 4.5:1 to about 5:1. In some embodiments, a subjectformulation comprises EPA and DHA in a ratio of 2.5:1 EPA:DHA.

The amount of vitamin E present in a subject formulation can beexpressed in units (International Units, or IU), or in milligrams. Inthe past, the U.S. Dietary Reference Intake (DRI) Recommended DietaryAllowances (RDA) of vitamin E were expressed in Units. This “units” termhas been replaced in recent years by α-tocopherol equivalents (“α-TE”)or milligrams. One Unit is equivalent to 1 mg of dl-α-tocopherol acetateor 0.6 mg d-α-tocopherol. Throughout this specification, amounts ofvitamin E are given in mg.

The α-tocopherol and γ-tocopherol isoforms of vitamin E can be presentin a subject formulation in an amount of from about 100 mg to about10,000 mg, e.g., from about 100 mg to about 200 mg, from about 200 mg toabout 300 mg, from about 300 mg to about 400 mg, from about 400 mg toabout 500 mg, from about 500 mg to about 600 mg, from about 600 mg toabout 700 mg, from about 700 mg to about 800 mg, from about 800 mg toabout 900 mg, from about 900 mg to about 1000 mg, from about 1000 mg toabout 2000 mg, from about 2000 mg to about 3000 mg, from about 3000 mgto about 3000 mg, from about 4000 mg to about 5000 mg, from about 5000mg to about 6000 mg, from about 6000 mg to about 7000 mg, from about7000 mg to about 8000 mg, from about 8000 mg to about 9000 mg, or fromabout 9000 mg to about 10,000 mg per unit dose, where the amounts givenare for individual isoforms of vitamin E. In some embodiments, theα-tocopherol is d-α-tocopherol.

In some embodiments, a subject formulation includes α-tocopherol in anamount of from about 500 mg to about 3000 mg per unit dose, e.g., in anamount of from about 500 mg, from about 500 mg to about 600 mg, fromabout 600 mg to about 700 mg, from about 700 mg to about 800 mg, fromabout 800 mg to about 900 mg, from about 900 mg to about 1000 mg, fromabout 1000 mg to about 2000 mg, or from about 2000 mg to about 3000 mgper unit dose; and γ-tocopherol in an amount of from about 200 mg toabout 1000 mg per unit dose, e.g., in an amount of from about 200 mg toabout 300 mg, from about 300 mg to about 400 mg, from about 400 mg toabout 500 mg, from about 500 mg to about 600 mg, from about 600 mg toabout 700 mg, from about 700 mg to about 800 mg, from about 800 mg toabout 900 mg, or from about 900 mg to about 1000 mg per unit dose. Insome of these embodiments, the formulation does not include vitamin Eisoforms other than α-tocopherol and γ-tocopherol. In other embodiments,the formulation includes, in addition to α-tocopherol and γ-tocopherol,at least one additional vitamin E isoform.

In some embodiments, a subject formulation includes α-tocopherol andγ-tocopherol; and does not include other vitamin E isoforms. In otherembodiments, a subject formulation includes α-tocopherol andγ-tocopherol; and one or more additional isoforms of vitamin E. Where asubject formulation includes α-tocopherol and γ-tocopherol, theα-tocopherol can be present in an amount of from about 500 mg to about10,000 mg, e.g., from about 500 mg to about 1000 mg, from about 1000 mgto about 2000 mg, from about 2000 mg to about 3000 mg, from about 3000mg to about 3000 mg, from about 4000 mg to about 5000 mg, from about5000 mg to about 6000 mg, from about 6000 mg to about 7000 mg, fromabout 7000 mg to about 8000 mg, from about 8000 mg to about 9000 mg, orfrom about 9000 mg to about 10,000 mg; and the γ-tocopherol can bepresent in an amount of from about 100 mg to about 1000 mg, e.g., fromabout 100 mg to about 200 mg, from about 200 mg to about 300 mg, fromabout 300 mg to about 400 mg, from about 400 mg to about 500 mg, fromabout 500 mg to about 600 mg, from about 600 mg to about 700 mg, fromabout 700 mg to about 800 mg, from about 800 mg to about 900 mg, or fromabout 900 mg to about 1000 mg per unit dose.

Other forms of vitamin E (e.g., β-tocopherol, δ-tocopherol,α-tocotrienol, β-tocotrienol, δ-tocotrienol, and γ-tocotrienol), ifpresent in a subject formulation, can be present in an amount of fromabout 5 mg to about 2000 mg, e.g., from about 5 mg to about 10 mg, fromabout 10 mg to about 25 mg, from about 25 mg to about 50 mg, from about50 mg to about 75 mg, from about 75 mg to about 100 mg, from about 100mg to about 125 mg, from about 125 mg to about 150 mg, from about 150 mgto about 175 mg, or from about 175 mg to about 200 mg, from about 200 mgto about 250 mg, from about 250 mg to about 500 mg, from about 500 mg toabout 750 mg, from about 750 mg to about 1000 mg, from about 1000 mg toabout 1250 mg, from about 1250 mg to about 1500 mg, from about 1500 mgto about 1750 mg, or from about 1750 mg to about 2000 mg per unit dose,where the amounts given are for the individual isoforms of vitamin E.

Where a subject formulation comprises one or more omega-6 fatty acids,the omega-6 fatty acid can be present in the formulation in an amount offrom about 50 mg to about 500 mg, e.g., from about 50 mg to about 75 mg,from about 75 mg to about 100 mg, from about 100 mg to about 150 mg,from about 150 mg to about 200 mg, from about 200 mg to about 250 mg,from about 250 mg to about 300 mg, from about 300 mg to about 350 mg,from about 350 mg to about 400 mg, from about 400 mg to about 450 mg, orfrom about 450 mg to about 500 mg per unit dose.

Where a subject formulation comprises one or more omega-9 fatty acids,the omega-9 fatty acid can be present in a subject formulation in anamount of from about 50 mg to about 500 mg, e.g., from about 50 mg toabout 75 mg, from about 75 mg to about 100 mg, from about 100 mg toabout 150 mg, from about 150 mg to about 200 mg, from about 200 mg toabout 250 mg, from about 250 mg to about 300 mg, from about 300 mg toabout 350 mg, from about 350 mg to about 400 mg, from about 400 mg toabout 450 mg, or from about 450 mg to about 500 mg per unit dose.

Where a subject formulation comprises α-lipoic acid, the α-lipoic acidcan be present in a subject formulation in an amount of from about 50 mgto about 1000 mg, e.g., from about 50 mg to about 75 mg, from about 75mg to about 100 mg, from about 100 mg to about 150 mg, from about 150 mgto about 200 mg, from about 200 mg to about 250 mg, from about 250 mg toabout 300 mg, from about 300 mg to about 350 mg, from about 350 mg toabout 400 mg, from about 400 mg to about 450 mg, from about 450 mg toabout 500 mg, from about 500 mg to about 600 mg, from about 600 mg toabout 700 mg, from about 700 mg to about 800 mg, from about 800 mg toabout 900 mg, or from about 900 mg to about 1000 mg per unit dose.

Where a subject formulation comprises carnitine, the carnitine can bepresent in a subject formulation in an amount of from about 150 mg toabout 3000 mg, e.g., from about 150 mg to about 200 mg, from about 200mg to about 250 mg, from about 250 mg to about 300 mg, from about 300 mgto about 350 mg, from about 350 mg to about 400 mg, from about 400 mg toabout 450 mg, from about 450 mg to about 500 mg, from about 500 mg toabout 600 mg, from about 600 mg to about 700 mg, from about 700 mg toabout 800 mg, from about 800 mg to about 900 mg, from about 900 mg toabout 1000 mg, from about 1000 mg to about 1250 mg, from about 1250 mgto about 1500 mg, from about 1500 mg to about 1750 mg, from about 1750mg to about 2000 mg, from about 2000 mg to about 2250 mg, from about2250 mg to about 2500 mg, from about 2500 mg to about 2750 mg, or fromabout 2750 mg to about 3000 mg per unit dose.

Additional Components

In some embodiments, a subject formulation includes, in addition to aPUFA and at least one vitamin E isoform, one or more additionalcomponents, e.g., one or more of: a) vitamin C (ascorbic acid); b) zinc;c) vitamin K; d) phosphocholine; e) coenzyme-Q; f) selenium; g) vitaminA; h) thiamin (vitamin B₁); i) riboflavin (vitamin B₂); j) pyridoxine,pyridoxamine, pyridoxine (collectively known as vitamin B₆); k)cyanocobalamin (vitamin B₁₂); l) vitamin D; m) arginine; n) calcium; o)magnesium; p) biotin (vitamin B₇); q) folic acid (vitamin B₉); r)pantothenic acid (vitamin B₅); s) tetrahydrobiopterin; and t) niacin orniacinamide (vitamin B₃). In some embodiments, a subject formulationincludes a PUFA; at least one vitamin E isoform; and one, two, three,four, five, six, seven, eight, nine, ten, 11, 12, 13, 14, 15, 16, 17,18, 19, or all twenty of: a) vitamin C (ascorbic acid); b) zinc; c)vitamin K; d) phosphocholine; e) coenzyme-Q; f) selenium; g) vitamin A;h) thiamin (vitamin B₁); i) riboflavin (vitamin B₂); j) pyridoxine,pyridoxamine, pyridoxine (collectively known as vitamin B₆); k)cyanocobalamin (vitamin B₁₂); l) vitamin D; m) arginine; n) calcium; o)magnesium; p) biotin (vitamin B₇); q) folic acid (vitamin B₉); r)pantothenic acid (vitamin B₅); s) tetrahydrobiopterin; and t) niacin orniacinamide (vitamin B₃). In some embodiments, a subject formulationincludes one or more amino acids. In some embodiments, a subjectformulation includes an anti-fungal agent (e.g., imidozoles andtriazoles, nystatin, amphotericin B etc). In some embodiments, a subjectformulation includes an anti-inflammatory agent. In some embodiments, asubject formulation includes an agent that reduces oxidative stress.

In some embodiments, a subject formulation includes a PUFA; at least onevitamin E isoform; and a pancreatic enzyme. In some embodiments, asubject formulation includes a PUFA; at least one vitamin E isoform; anda leukotriene inhibitor. In some embodiments, a subject formulationincludes a PUFA; at least one vitamin E isoform; and a mast cellstabilizer (e.g., cromolyn). In other embodiments, a subject formulationincludes a PUFA; at least one vitamin E isoform; a pancreatic enzyme;and a leukotriene inhibitor. For example, in some embodiments, a subjectformulation includes an omega-3 fatty acid (e.g., EPA; DHA; or acombination of EPA and DHA, as described above; vitamin E (e.g.,α-tocopherol and γ-tocopherol); α-lipoic acid; carnitine; a pancreaticenzyme; and a leukotriene inhibitor. Pancreatic enzymes and leukotrieneinhibitors are described in more detail below.

Any minerals in a subject formulation can be present in salt form. Suchsalts can be any of the well known salts including carbonate, oxide,hydroxide, chloride, sulfate, phosphate, gluconate, lactate, acetate,fumarate, citrate, malate, amino acids, and the like for the cationicminerals and sodium, potassium, calcium, magnesium, and the like for theanionic minerals.

a) Vitamin C

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin C. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin C. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin C.

“Vitamin C” includes all forms of ascorbic acid, such as L-ascorbicacid, D-ascorbic acid, DL-ascorbic acid, D-araboascorbic acid,dehydroascorbic acid, xyloascorbic acid, esters of ascorbic acid, saltsof ascorbic acid, and the like.

Vitamin C can be present in a subject formulation in an amount of fromabout 80 mg to about 1000 mg, from about 80 mg to about 100 mg, fromabout 100 mg to about 200 mg, from about 200 mg to about 250 mg, fromabout 250 mg to about 300 mg, from about 300 mg to about 400 mg, fromabout 400 mg to about 500 mg, from about 500 mg to about 600 mg, fromabout 600 mg to about 700 mg, from about 700 mg to about 800 mg, fromabout 800 mg to about 900 mg, or from about 900 to about 1000 mg totaldaily dose or per unit dose.

b) Zinc

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and zinc. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and zinc. In otherembodiments, a subject formulation includes a PUFA (e.g., an omega-3fatty acid and/or an omega-6 fatty acid); at least one isoform ofvitamin E; α-lipoic acid; and zinc.

Zinc can be present in a subject formulation in the form of zincgluconate, zinc sulfate, zinc chloride, or any salt of zinc. Zinc can bepresent in a subject formulation in an amount of from about 5 mg toabout 50 mg, from about 5 mg to about 10 mg, from about 10 mg to about15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg,from about 25 mg to about 30 mg, from about 30 mg to about 40 mg, orfrom about 40 mg to about 50 mg total daily dose or per unit dose.

c) Vitamin K

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin K. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin K. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin K.

“Vitamin K” includes Vitamin K₁ (phytonadione, phylloquinone,phytonactone); Vitamin K₂ (menaquinones, e.g., menaquinone-4,menaquinone-7, etc.); Vitamin K₃ (menadione; or2-methyl-1,4-naphthoquinone); a salt of a vitamin K; and a derivative ofa vitamin K. In some embodiments, a subject formulation includes vitaminK₁. In other embodiments, a subject formulation includes vitamin K₂. Inother embodiments, a subject formulation includes vitamin K₁ and vitaminK₂.

Vitamin K₁ can be present in a subject formulation in an amount of fromabout 100 μg to about 10 mg (total daily dose), e.g., from about 100 μgto about 500 μg, from about 500 μg to about 1 mg, from about 1 mg toabout 2.5 mg, from about 2.5 mg to about 5 mg, from about 5 mg to about7.5 mg, or from about 7.5 mg to about 10 mg.

Vitamin K₂ can be present in a subject formulation in an amount of fromabout 100 μg to about 2 mg (total daily dose), e.g., from about 100 μgto about 250 μg, from about 250 μg to about 500 μg, from about 500 μg toabout 750 μg, from about 750 μg to about 1 mg, from about 1 mg to about1.25 mg, from about 1.25 mg to about 1.5 mg, from about 1.5 mg to about1.75 mg, or from about 1.75 mg to about 2 mg.

In some embodiments, a subject formulation includes vitamin K₁. In otherembodiments, a subject formulation includes vitamin K₂. In otherembodiments, a subject formulation includes both vitamin K₁ and vitaminK₂.

d) Phosphocholine

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and phosphocholine. In some embodiments, a subjectformulation includes a PUFA (e.g., an omega-3 fatty acid and/or anomega-6 fatty acid); at least one isoform of vitamin E; carnitine; andphosphocholine. In other embodiments, a subject formulation includes aPUFA (e.g., an omega-3 fatty acid and/or an omega-6 fatty acid); atleast one isoform of vitamin E; α-lipoic acid; and phosphocholine.“Phosphocholine” includes phosphatidylcholine derivatives ofphosphocholine, e.g., polyenylphosphatidylcholine.

Phosphocholine (or a phosphatidylcholine such aspolyenylphosphatidylcholine) can be present in a subject formulation inan amount of from about 500 mg to about 5000 mg, e.g., from about 500 mgto about 1000 mg, from about 1000 mg to about 2000 mg, from about 2000mg to about 3000 mg, from about 3000 mg to about 4000 mg, or from about4000 mg to about 5000 mg total daily dose or per unit dose.

e) Coenzyme Q

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and Coenzyme Q. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and Coenzyme Q. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and Coenzyme Q.

“Coenzyme Q” (CoQ; 2,3-dimethoxy-5-methyl-6-polyprenyl-1,4-benzoquinone;also known as ubiquinone) refers to a group of lipid solublebenzoquinones involved in electron transport in mitochondrialpreparations, e.g., in the oxidation of succinate or reduced nicotineadenine dinucleotide (NADH) via the cytochrome system. CoQ includesCoQ_(n), where n is an integer from 1 to 12, and where n indicates thenumber of isoprenoid units. CoQ includes, e.g., CoQ₇₋₁₀, i.e. CoQ₇(ubiquinone-7), CoQ₉ (ubiquinone-9), CoQ₁₀ (ubidecarenone), and mixturesof the foregoing. In some embodiments, the CoQ is ubidecarenone.

CoQ can be present in a subject formulation in an amount of from about 4mg to about 250 mg, from about 4 mg to about 10 mg, from about 10 mg toabout 25 mg, from about 25 mg to about 50 mg, from about 50 mg to about75 mg, from about 75 mg to about 100 mg, from about 100 mg to about 125mg, from about 125 mg to about 150 mg, from about 150 mg to about 200mg, or from about 200 mg to about 250 mg total daily dose or per unitdose.

f) Selenium

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and selenium. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and selenium. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and selenium.

Selenium can be present in the form of sodium selenate, sodium selenite,selenomethionine, and the like. Selenium can be present in a subjectformulation in an amount of from about 20 μg to about 500 kg, from about20 kg to about 30 kg, from about 30 kg to about 40 kg, from about 40 kgto about 50 kg, from about 50 kg to about 75 kg, from about 75 kg toabout 100 kg, from about 100 kg to about 150 kg, from about 150 kg toabout 200 kg, from about 200 kg to about 250 kg, from about 250 kg toabout 300 kg, from about 300 kg to about 350 kg, from about 350 kg toabout 400 kg, or from about 400 kg to about 500 kg total daily dose.

g) Vitamin A

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin A. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin A. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin A.

“Vitamin A” includes retinol, retinal (retinaldehyde), and retinoicacid; nor-retinoids; retro-retinoids; seco-retinoids (e.g.,1,6-seco-1,2-didehydroretinol); substituted retinoids (e.g.,5,6-Epoxy-5,6-dihydroretinol; ethyl 12-fluororetinoate; etc.); andanalogs such as 3-hydroxyretinol, 3-hydroxyretinoic acid,3-hydroxyretinal, 4-oxoretinol, 4-oxoretinoic acid, 4-oxoretinal,3,4-didehydroretinol (vitamin A₂), 3,4-didehydroretinoic acid,3,4-didehydroretinal, 4,5-didehydro-5,6-dihydroretinol, acycloretinol,acycloretinoic acid, and acycloretinal; and esters of vitamin A, e.g.,an acetate ester, a succinate ester, a palmitate ester, etc.

Vitamin A can be present in a subject formulation in an amount of fromabout 200 IU to about 10,000 IU, e.g., 200 IU to about 250 IU, fromabout 250 IU to about 500 IU, from about 500 IU to about 1000 IU, fromabout 1000 IU to about 2000 IU, from about 2000 IU to about 3000 IU,from about 3000 IU to about 4000 IU, from about 4000 IU to about 5000IU, from about 5000 IU to about 7500 IU, or from about 7500 IU to about10,000 IU total daily dose.

h) Vitamin B₁

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₁. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₁. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₁.

“Vitamin B₁” includes thiamine (also referred to as “thiamin”); thehydrochloride and nitrate salts of thiamine; and thiamine alkyldisulfides such as thiamine propyldisulfide, thiamine tetrahydrofurfuryldisulfide, and thiamine o-benzoyl disulfide; neopyrithiamine;oxyneopyrithiamine; and the like.

Vitamin B₁ can be present in a subject formulation in an amount of fromabout 0.05 mg to about 15 mg, from about 0.05 mg to about 0.1 mg, fromabout 0.1 mg to about 1.0 mg, from about 1.0 mg to about 1.5 mg, fromabout 1.5 mg to about 2.0 mg, from about 2.0 mg to about 2.5 mg, fromabout 2.5 mg to about 5 mg, from about 5 mg to about 7.5 mg, from about7.5 mg to about 10 mg, from about 10 mg to about 12.5 mg, or from about12.5 mg to about 15 mg total daily dose or per unit dose.

i) Vitamin B₂

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₂. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₂. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₂.

“Vitamin B₂” includes riboflavin; crystalline riboflavin coenzyme formsof riboflavin such as flavin adenine dinucleotide, flavin adeninemononucleotide, riboflavin 5-phosphate and their salts. Vitamin B₂ canbe present in a subject formulation in an amount of from about 0.05 mgto about 15 mg, from about 0.05 mg to about 0.1 mg, from about 0.1 mg toabout 1.0 mg, from about 1.0 mg to about 1.5 mg, from about 1.5 mg toabout 2.0 mg, from about 2.0 mg to about 2.5 mg, from about 2.5 mg toabout 5 mg, from about 5 mg to about 7.5 mg, from about 7.5 mg to about10 mg, from about 10 mg to about 12.5 mg, or from about 12.5 mg to about15 mg total daily dose or per unit dose.

j) Vitamin B₆

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₆. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₆. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₆.

“Vitamin B₆” includes pyridoxine, pyridoxal, pyridoxamine; andhydrochloride salts or 5-phosphates of pyridoxine, pyridoxamine, orpyridoxal. For example, pyridoxine hydrochloride can be included in asubject formulation.

Vitamin B₆ can be present in a subject formulation in an amount of fromabout 2 mg to about 250 mg, from about 2 mg to about 5 mg, from about 5mg to about 10 mg, from about 10 mg to about 25 mg, from about 25 mg toabout 50 mg, from about 50 mg to about 75 mg, from about 75 mg to about100 mg, from about 100 mg to about 125 mg, from about 125 mg to about150 mg, from about 150 mg to about 175 mg, from about 175 mg to about200 mg, from about 200 mg to about 225 mg, or from about 225 mg to about250 mg total daily dose or per unit dose.

k) Vitamin B 12

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₁₂. In some embodiments, a subjectformulation includes a PUFA (e.g., an omega-3 fatty acid and/or anomega-6 fatty acid); at least one isoform of vitamin E; carnitine; andvitamin B₁₂. In other embodiments, a subject formulation includes a PUFA(e.g., an omega-3 fatty acid and/or an omega-6 fatty acid); at least oneisoform of vitamin E; α-lipoic acid; and vitamin B₁₂.

“Vitamin B₁₂” refers to vitamin B₁₂ (cyanocobalamin) and itspharmaceutical derivatives, such as hydroxocobalamin,cyano-10-chlorocobalamin, aquocobalamin perchlorate,aquo-10-chlorocobalamin perchlorate, azidocobalamin, chlorocobalamin,cobalamin, methylcobalamin, adenosylcobalamin, and hydroxocobalamin.

Vitamin B₁₂ can be present in a subject formulation in an amount of fromabout 2 μg to about 1000 μg, 2 μg to about 10 μg, from about 10 μg toabout 25 μg, from about 25 μg to about 50 μg, from about 50 μg to about100 μg, from about 100 μg to about 200 μg, from about 200 μg to about300 μg, from about 300 μg to about 400 μg, from about 400 μg to about500 μg, from about 500 μg to about 600 μg, from about 600 μg to about700 μg, from about 700 μg to about 800 μg, from about 800 μg to about900 μg, or from about 900 μg to about 1000 μg total daily dose or perunit dose.

l) Vitamin D

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin D. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin D. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin D.

“Vitamin D” includes, e.g., cholecalciferol (D3), ergocalciferol (D2),and biologically active metabolites and precursors such as, e.g.,1-α-hydroxy Vitamin D, 25-hydroxy Vitamin D, 1,25-dihydroxy Vitamin D,and the like.

Vitamin D can be present in a subject formulation in an amount of fromabout 200 IU to about 800 IU, from about 200 IU to about 300 IU, fromabout 300 IU to about 400 IU, from about 400 IU to about 500 IU, fromabout 500 IU to about 600 IU, from about 600 IU to about 700 IU, or fromabout 700 IU to about 800 IU total daily dose or per unit dose.

m) Arginine

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and arginine. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and arginine. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and arginine.

“Arginine” includes L-arginine, and arginine analogs such as methylatedderivatives; and the like. Arginine (e.g., L-arginine) can be present ina subject formulation in an amount of from about 75 mg to about 9000 mg,from about 75 mg to about 100 mg, from about 100 mg to about 250 mg,from about 250 mg to about 500 mg, from about 500 mg to about 750 mg,from about 750 mg to about 1000 mg, from about 1000 mg to about 1500 mg,from about 1500 mg to about 2000 mg, from about 2000 mg to about 3000mg, from about 3000 mg to about 4000 mg, from about 4000 mg to about5000 mg, from about 5000 mg to about 6000 mg, from about 6000 mg toabout 7000 mg, from about 7000 mg to about 8000 mg, or from about 8000mg to about 9000 mg per unit dose.

n) Calcium

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and calcium. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and calcium. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and calcium.

Calcium can be present in a subject formulation in the form of calciumcarbonate, calcium citrate, calcium glubionate, calcium gluconate,calcium lactate, dibasic calcium phosphate, tribasic calcium phosphate,calcium acetate, and the like. Calcium can be present in a subjectformulation in an amount of from about 40 mg to about 2000 mg, fromabout 40 mg to about 100 mg, from about 100 mg to about 250 mg, fromabout 250 mg to about 500 mg, from about 500 mg to about 750 mg, fromabout 750 mg to about 1000 mg, from about 1000 mg to about 1500 mg, orfrom about 1500 mg to about 2000 mg total daily dose or per unit dose.

o) Magnesium

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and magnesium. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and magnesium. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and magnesium.

Magnesium can be present in a subject formulation as magnesium oxide,magnesium citrate, magnesium chloride, magnesium gluceptate, magnesiumhydroxide, magnesium gluconate, magnesium lactate, magnesium pidolate,magnesium sulfate, and the like. Magnesium can be present in a subjectformulation in an amount of from about 50 mg to about 450 mg, from about50 mg to about 75 mg, from about 75 mg to about 100 mg, from about 100mg to about 150 mg, from about 150 mg to about 200 mg, from about 200 mgto about 250 mg, from about 250 mg to about 300 mg, from about 300 mg toabout 350 mg, from about 350 mg to about 400 mg, or from about 400 mg toabout 450 mg total daily dose or per unit dose.

p) Vitamin B₇

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₇. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₇. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₇.

“Vitamin B₇” includes biotin, oxybiotin, biocytin, biotinol,D-homobiotin, D-norbiotin, dethiobiotin, biotin sulfone, biotin diaminesulfate, and the like. Vitamin B₇ can be present in a subjectformulation in an amount of from about 10 Ctg to about 800 μg, fromabout 10 μg to about 25 μg, from about 25 μg to about 50 μg, from about50 μg to about 100 μg, from about 100 μg to about 200 μg, from about 200μg to about 300 μg, from about 300 μg to about 400 μg, from about 400 μgto about 500 μg, from about 500 μg to about 600 μg, from about 600 μg toabout 700 μg, or from about 700 μg to about 800 μg total daily dose orper unit dose.

q) Vitamin B₉

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₉. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₉. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₉.

“Folic acid” (also known as pteroyl-L-glutamic acid; or vitamin B₉)includes folic acid and any chemical derivative of folic acid thatfunction equivalently to folic acid, such as mono and polyglutamylfolates, dihydro and tetrahydro folates, methyl and formyl folates, andany isomer of a folate, e.g., an isomer of a reduced folate. Thus, e.g.,“folic acid” includes dihydrofolic acid, tetrahydrofolic acid,5-formyltetrahydrofolic acid, 10-formyltetrahydrofolic acid, 5-10methylenetetrahydrofolic acid, 5-10 methenyltetrahydrofolic acid,5-methyltetrahydrofolic acid, and derivatives of any of the foregoing;and a natural isomer of reduced folate, such as (6S)-tetrahydrofolicacid, 5-methyl-(6S)-tetrahydrofolic acid, 5-formyl-(6S)-tetrahydrofolicacid, 10-formyl-(6R)-tetrahydrofolic acid,5,10-methylene-(6R)-tetrahydrofolic acid,5,10-methenyl-(6R)-tetrahydrofolic acid,5-formimino-(6S)-tetrahydrofolic acid, and polyglutamyl derivatives ofany of the foregoing. Derivatives of folic acid include the precursors(pro-vitamins), metabolites, derivatives, and conjugates of the parentcompound, any of which may be either naturally occurring or synthetic;as well as the salts of the compounds. “Folic acid” includes naturallyoccurring and synthetic forms of folic acid.

Folic acid can be present in a subject formulation in an amount of fromabout 200 μg to about 1000 μg, e.g., from about 200 μg to about 400 μg,from about 400 μg to about 500 μg, from about 500 μg to about 600 μg,from about 600 μg to about 700 μg, from about 700 μg to about 800 μg,from about 800 μg to about 900 μg, or from about 900 μg to about 1000 μgtotal daily dose or per unit dose.

r) Vitamin B₅

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₅. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₅. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₅.

“Vitamin B₅” is also referred to as pantothenate or pantothenic acid,and encompasses salts such as calcium pantothenate; pantothenol; andpanthenol. Vitamin B₅ can be present in a subject formulation in anamount of from about 0.4 mg to about 800 mg, from about 0.4 mg to about1 mg, from about 1 mg to about 10 mg, from about 10 mg to about 25 mg,from about 25 mg to about 50 mg, from about 50 mg to about 100 mg, fromabout 100 mg to about 200 mg, from about 200 mg to about 300 mg, fromabout 300 mg to about 400 mg, from about 400 mg to about 500 mg, fromabout 500 mg to about 600 mg, from about 600 mg to about 700 mg, or fromabout 700 mg to about 800 mg total daily dose or per unit dose.

s) Tetrahydrobiopterin

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and tetrahydrobiopterin. In some embodiments, a subjectformulation includes a PUFA (e.g., an omega-3 fatty acid and/or anomega-6 fatty acid); at least one isoform of vitamin E; carnitine; andtetrahydrobiopterin. In other embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; α-lipoic acid; andtetrahydrobiopterin. In other embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; α-lipoic acid; arginine; andtetrahydrobiopterin. In other embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; α-lipoic acid; carnitine; andtetrahydrobiopterin. In other embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; α-lipoic acid; arginine;carnitine; and tetrahydrobiopterin.

“Tetrahydrobiopterin” includes 5,6,7,8-tetrahydrobiopterin and saltsthereof; L-erythro-5,6,7,8-tetrahydrobiopterin and salts thereof; andthe like. Tetrahydrobiopterin can be present in a subject formulation inan amount of from about 20 mg to about 3000 mg, from about 20 mg toabout 50 mg, from about 50 mg to about 100 mg, from about 100 mg toabout 250 mg, from about 250 mg to about 500 mg, from about 500 mg toabout 750 mg, from about 750 mg to about 1000 mg, from about 1000 mg toabout 1500 mg, from about 1500 mg to about 2000 mg, from about 2000 mgto about 2500 mg, or from about 2500 mg to about 3000 mg.

t) Vitamin B₃

In some embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; and vitamin B₃. In some embodiments, a subject formulationincludes a PUFA (e.g., an omega-3 fatty acid and/or an omega-6 fattyacid); at least one isoform of vitamin E; carnitine; and vitamin B₃. Inother embodiments, a subject formulation includes a PUFA (e.g., anomega-3 fatty acid and/or an omega-6 fatty acid); at least one isoformof vitamin E; α-lipoic acid; and vitamin B₃.

“Vitamin B₃” includes niacin, niacinamide, nicotinic acid, nicotinamide(niacinamide), the coenzyme forms of niacin such as nicotinamide adeninedinucleotide, and nicotinamide adenine dinucleotide phosphate. VitaminB₃ can be present in a subject formulation in an amount of from about0.5 mg to about 200 mg, from about 0.5 mg to about 1 mg, from about 1 mgto about 10 mg, from about 10 mg to about 25 mg, from about 25 mg toabout 50 mg, from about 50 mg to about 75 mg, from about 75 mg to about100 mg, from about 100 mg to about 150 mg, or from about 150 mg to about200 mg.

Pancreatic Enzymes

As noted above, in some embodiments, a subject formulation comprises, inaddition to a PUFA and at least one vitamin E isoform, a pancreaticenzyme.

Pancreatic enzymes include proteases, lipases, amylases, and nucleases.The term “pancreatic enzyme,” as used herein, refers to any enzyme thatprovides a function (catalytic activity) of an enzyme produced by ahuman pancreas. For example, “pancreatic lipase” refers to any enzymethat provides a function of a lipase produced by a human pancreas.

In some embodiments, a subject formulation includes a lipase, e.g., apancreatic lipase. In other embodiments, a subject formulation includesa pancreatic protease (e.g., trypsin, trypsinogen, chymotrypsin,chymotrypsinogen), and a pancreatic lipase. In other embodiments, asubject formation includes a pancreatic protease, a pancreatic lipase,and a pancreatic amylase.

In some embodiments, the enzyme is an inactive proenzyme (e.g.,trypsinogen, chymotrysinogen). A pancreatic enzyme to be included in asubject formulation can be a naturally-occurring enzyme, a recombinantenzyme, or a synthetic enzyme; and can be from any of a variety ofsources, e.g., a mammal, a fungus, a plant, etc. For example, fungalenzymes, and formulations comprising same, are described in, e.g., U.S.Pat. No. 6,051,220. In some embodiments, the enzyme is acid stable,e.g., is stable at a pH range of from about 2.8 to about 9. In someembodiments, the enzyme is in a microencapsulated and enteric coatedformulation. Examples of such formulations include, e.g., Cotazym-S,Creon, Pancrease, Pancrease MT-16, Ultrase MT-20, and Zymase. Othersuitable formulations include, e.g., a formulation as described in U.S.Pat. No. 5,750,104.

The amount of pancreatic enzyme present in a subject formulation canvary, according to need, and can be in a range of from about 2000International Units (IU) to 40,000 IU per unit dose, e.g., from about2000 IU to about 5000 IU, from about 5000 IU to about 7,500 IU, fromabout 7,500 IU to about 10,000 IU, from about 10,000 IU to about 15,000IU, from about 15,000 IU to about 20,000 IU, from about 20,000 IU toabout 30,000 IU, or from about 30,000 IU to about 40,000 IU per unitdose.

Leukotriene Inhibitors

As noted above, in some embodiments, a subject formulation comprises, inaddition to a PUFA and at least one vitamin E isoform, a leukotrieneinhibitor. Suitable leukotriene inhibitors include leukotriene receptorantagonists and leukotriene synthesis inhibitors. Exemplary leukotrienereceptor antagonists include, e.g., zafirlukast (Accolate), montelukast(Singulair), pranlukast, iralukast, pobilukast and SKB-106,203.Leukotriene synthesis inhibitors include inhibitors of 5-lipoxygenaseactivity, where an exemplary 5-lipoxygenase inhibitor is zileuton(Zyflo). Suitable 5-lipoxygenase inhibitors include those described in,e.g., U.S. Pat. Nos. 5,364,877, 5,302,603, 5,234,950, 5,098,932 and5,354,865. In some embodiments, a 5-lipoxygenase inhibitor also inhibitscyclooxygenase-2. In other embodiments, a 5-lipoxygenase inhibitor is aselective 5-lipoxygenase inhibitor, e.g., the inhibitor does notsubstantially inhibit enzymes other than 5-lipoxygenase, e.g., theinhibitor does not substantially inhibit a cyclooxygenase.

The amount of a leukotriene inhibitor that is included in a subjectformulation can vary, depending on factors such as the age and/or weightof the individual to whom the formulation is administered, the severityof symptoms, etc. The amount of a leukotriene inhibitor that is includedin a subject formulation can range from about 2 mg to about 100 mg,e.g., from about 2 mg to about 5 mg, from about 5 mg to about 10 mg,from about 10 mg to about 20 mg, from about 20 mg to about 30 mg, fromabout 30 mg to about 40 mg, from about 40 mg to about 50 mg, from about50 mg to about 60 mg, from about 60 mg to about 70 mg, from about 70 mgto about 80 mg, from about 80 mg to about 90 mg, or from about 90 mg toabout 100 mg per unit dose.

Amino Acids

In some embodiments, a subject formulation includes a PUFA; at least onevitamin E isoform; and one or more amino acids. For example, in someembodiments, a subject formulation includes arginine, glutamine, or botharginine and glutamine. If arginine and/or glutamine is present in asubject formulation, the arginine and/or glutamine is present in anamount of from about 500 mg to about 10 g per unit dose, e.g., fromabout 500 mg to about 750 mg, from about 750 mg to about 1 g, from about1 g to about 2.5 g, from about 2.5 g to about 5 g, from about 5 g toabout 7.5 g, or from about 7.5 g to about 10 g. For example, where asubject formulation includes a PUFA; at least one vitamin E isoform; andglutamine, the glutamine is present in an amount of from about 500 mg toabout 10 g per unit dose, e.g., from about 500 mg to about 750 mg, fromabout 750 mg to about 1 g, from about 1 g to about 2.5 g, from about 2.5g to about 5 g, from about 5 g to about 7.5 g, or from about 7.5 g toabout 10 g. As another example, where a subject formulation includes aPUFA; at least one vitamin E isoform; and arginine, the arginine ispresent in an amount of from about 500 mg to about 10 g per unit dose,e.g., from about 500 mg to about 750 mg, from about 750 mg to about 1 g,from about 1 g to about 2.5 g, from about 2.5 g to about 5 g, from about5 g to about 7.5 g, or from about 7.5 g to about 10 g. As anotherexample, where a subject formulation includes a PUFA; at least onevitamin E isoform; and both arginine and glutamine, the arginine andglutamine are each present in an amount of from about 500 mg to about 10g per unit dose, e.g., from about 500 mg to about 750 mg, from about 750mg to about 1 g, from about 1 g to about 2.5 g, from about 2.5 g toabout 5 g, from about 5 g to about 7.5 g, or from about 7.5 g to about10 g.

Anti-Fungal Agents

As noted above, in some embodiments, a subject formulation comprises, inaddition to a PUFA and at least one vitamin E isoform, an anti-fungalagent. Suitable anti-fungal agents include, but are not limited to,nystatin, amphotericin B, clotrimazole, flucytosine, fluconazole,itraconazole, ketoconazole, and noxafil. Where the anti-fungal agent isnystatin, the nystatin can be present in a subject formulation in anamount of from about 100,000 Units (U) to about 800,000 U per unit dose,e.g., from about 100,000 U to about 200,000 U, from about 200,000 U toabout 400,000 U, from about 400,000 U to about 600,000 U, or from about600,000 U to about 800,000 U per unit dose. In some embodiments, asubject formulation comprises a PUFA, at least one vitamin E isoform(e.g., α-tocopherol and γ-tocopherol), carnitine, and nystatin.

Anti-Inflammatory Agents

As noted above, in some embodiments, a subject formulation comprises, inaddition to a PUFA and at least one vitamin E isoform, ananti-inflammatory agent. Suitable anti-inflammatory agents include, butare not limited to, steroidal anti-inflammatory agents, andnon-steroidal anti-inflammatory agents. Suitable steroidalanti-inflammatory agents include, but are not limited to,hydrocortisone, hydroxyltriamcinolone, alpha-methyl dexamethasone,dexamethasone-phosphate, beclomethasone dipropionate, clobetasolvalerate, desonide, desoxymethasone, desoxycorticosterone acetate,dexamethasone, dichlorisone, diflorasone diacetate, diflucortolonevalerate, fluadrenolone, fluclorolone acetonide, fludrocortisone,flumethasone pivalate, fluosinolone acetonide, fluocinonide, flucortinebutylester, fluocortolone, fluprednidene (fluprednylidene) acetate,flurandrenolone, halcinonide, hydrocortisone acetate, hydrocortisonebutyrate, methylprednisolone, triamcinolone acetonide, conisone,cortodoxone, flucetonide, fludrocortisone, difluorosone diacetate,fluradrenolone acetonide, medrysone, amcinafel, amcinafide,betamethasone and the balance of its esters, chloroprednisone,chlorprednisone acetate, clocortelone, clescinolone, dichlorisone,difluprednate, flucloronide, flunisolide, fluoromethalone, fluperolone,fluprednisolone, hydrocortisone valerate, hydrocortisonecyclopentylpropionate, hydrocortamate, meprednisone, paramethasone,prednisolone, prednisone, beclomethasone dipropionate, triamcinolone,and mixtures of two or more of the foregoing.

Suitable non-steroidal anti-inflammatory agents, include, but are notlimited to, 1) the oxicams, such as piroxicam, isoxicam, tenoxicam, andsudoxicam; 2) the salicylates, such as aspirin, disalcid, benorylate,trilisate, safapryn, solprin, diflunisal, and fendosal; 3) the aceticacid derivatives, such as diclofenac, fenclofenac, indomethacin,sulindac, tolmetin, isoxepac, furofenac, tiopinac, zidometacin,acematacin, fentiazac, zomepiract, clidanac, oxepinac, and felbinac; 4)the fenamates, such as mefenamic, meclofenamic, flufenamic, niflumic,and tolfenamic acids; 5) the propionic acid derivatives, such asibuprofen, naproxen, benoxaprofen, flurbiprofen, ketoprofen, fenoprofen,fenbufen, indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen,miroprofen, tioxaprofen, suprofen, alminoprofen, and tiaprofenic; and 6)the pyrazoles, such as phenylbutazone, oxyphenbutazone, feprazone,azapropazone, and trimethazone, mixtures of these non-steroidalanti-inflammatory agents may also be employed, as well as thepharmaceutically-acceptable salts and esters of these agents.

Anti-Oxidants

As noted above, in some embodiments, a subject formulation comprises, inaddition to a PUFA and at least one vitamin E isoform, an agent thatreduces oxidative stress, e.g., an anti-oxidant. Suitable anti-oxidantsinclude, but are not limited to, NXY-059 (Disufenton sodium);chain-breaking phenolic antioxidants (such as Vitamin E and butylatedhydroxytoluene [BHT]); phenyl-substituted nitrones; azulenyl-substitutednitrones; α-phenyl-N-tert-butyl nitrone; stilbazulenyl nitrone (STAZN;Becker et al. (2002) J. Am. Chem. Soc. 124:4678); a spin-trap agent suchas, e.g., N-t-butyl-a-phenylnitrone,3,5-dibromo-4-nitrosobenzenesulfonic acid, 5,5-dimethyl-1-pyrrolineN-oxide, 2-methyl-2-nitrosopropane, nitrosodisulfonic acid,a-(4-pyridyl-1-oxide)-N-t-butylnitrone, 3,3,5,5-tetramethylpyrrolineN-oxide, 2,4,6-tri-t-butylnitrosobenzene, PTIYO(4-phenyl-2,2,5,5-tetramethyl imidazolin-1-yloxy-5-oxide), tempol(4-hydroxy 2,2,6,6-tetramethylpiperidine-1-oxyl); and the like. Ananti-oxidant can be a nitrone anti-oxidant (e.g., STAZN), a polyphenolanti-oxidant, a flavonol anti-oxidant (e.g., baicalein), or aphenylpropanoid anti-oxidant (e.g., chlorogenic acid, fisetin, etc.).Also suitable is an anti-oxidant as described in U.S. Patent PublicationNo. 2007/0275932.

Additional Components

A subject formulation can include one or more additional componentsother than those listed above. Such additional components can include,e.g., pharmaceutically acceptable components such as lactose, glucose,sucrose, corn starch, potato starch, cellulose esters such as celluloseacetate, ethyl cellulose, magnesium stearate, calcium silicate,precipitated silica, talc, fatty acids such as stearic acid,microcrystalline cellulose, carnauba wax and the like. Diluents andother additives such as one or more pharmaceutically acceptable bindingagents, fillers, supports, thickening agents, taste-improving agents,coloring agents, preservatives, stabilizers, regulators, emulsifiers,flow agents, absorbents, and the like or mixtures thereof may be useddepending on the form of the composition employed.

Food-Grade Components

In some embodiments, a subject formulation further includes one or morefood-grade components. Suitable components include, but are not limitedto, mono- and disaccharides; carbohydrates; proteins; amino acids; fattyacids; lipids; stabilizers; preservatives; flavoring agents; coloringagents; sweeteners; antioxidants, chelators, and carriers; texturants;pH adjusters; emulsifiers; stabilizers; soy and soy-based components;milk base solids; edible fibers; and the like. The food component can beisolated from a natural source, or can be synthesized. All componentsare food-grade components fit for human consumption.

Examples of suitable monosaccharides include sorbitol, mannitol,erythrose, threose, ribose, arabinose, xylose, ribulose, glucose,galactose, mannose, fructose, and sorbose. Non-limiting examples ofsuitable disaccharides include sucrose, maltose, lactitol, maltitol,maltulose, and lactose.

Suitable carbohydrates include oligosaccharides, polysaccharides, and/orcarbohydrate derivatives. As used herein, the term “oligosaccharide”refers to a digestible linear molecule having from 3 to 9 monosaccharideunits, wherein the units are covalently connected via glycosidic bonds.As used herein, the term “polysaccharide” refers to a digestible (i.e.,capable of metabolism by the human body) macromolecule having greaterthan 9 monosaccharide units, wherein the units are covalently connectedvia glycosidic bonds. The polysaccharides may be linear chains orbranched. Carbohydrate derivatives, such as a polyhydric alcohol (e.g.,glycerol), may also be utilized as a complex carbohydrate herein. Asused herein, the term “digestible” in the context of carbohydratesrefers to carbohydrates that are capable of metabolism by enzymesproduced by the human body. Examples of non-digestible carbohydrates areresistant starches (e.g., raw corn starches) and retrograded amyloses(e.g., high amylose corn starches). Non-limiting examples of suitablecarbohydrates include raffinoses, stachyoses, maltotrioses,maltotetraoses, glycogens, amyloses, amylopectins, polydextroses, andmaltodextrins.

Suitable starches include natural starches, e.g., starches derived froma natural source, such as those obtained from various plant species.Examples of plant sources of starch include, but are not limited to,corn, waxy corn, wheat, rice, tapioca, potato, pea and other sourcesknown in the art. Suitable starches include modified starches. Modifiedstarches are known in the art and the term generally refers to starchthat has been physically or chemically altered to improve its functionalcharacteristics. Suitable modified starches include, but are not limitedto, pre-gelatinized starches, low viscosity starches (such as dextrins,acid-modified starches, oxidized starches and enzyme modified starches),derivatized starches, stabilized starches (such as starch esters andstarch ethers), cross-linked starches, starch sugars (such as glucosesyrup, dextrose and isoglucose) and starches that have been submitted toa combination of treatments (such as cross-linking and gelatinization)and mixtures thereof.

In some embodiments, a subject formulation that comprises one or morefood components is gluten free. In some embodiments, a subjectformulation that comprises one or more food components is casein free.In some embodiments, a subject formulation that comprises one or morefood components is gluten free and casein free. “Gluten free” means thata subject formulation contains substantially no gluten; or, if theformulation does contain gluten, the gluten is present in an amount thatdoes not induce an adverse reaction in an individual who is glutensensitive (e.g., allergic to gluten) or who is gluten intolerant.Similarly, “casein free” means that a subject formulation containssubstantially no gluten; or, if the formulation does contain gluten, thegluten is present in an amount that does not induce an adverse reactionin an individual who is casein sensitive (e.g., allergic to casein) orwho is casein intolerant.

Suitable fats include, but are not limited to, triglycerides, includingshort-chain (C₂-C₄) and long-chain triglycerides (C₁₆-C₂₂).

Suitable texturants (also referred to as soluble fibers) include, butare not limited to, pectin (high ester, low ester); carrageenan;alginate (e.g., alginic acid, sodium alginate, potassium alginate,calcium alginate); guar gum; locust bean gum; psyllium; xanthan gum; gumarabic; fructo-oligosaccharides; inulin; agar; a modified cellulose gum;and functional blends of two or more of the foregoing.

Suitable modified cellulose gums include, for example, methylcellulose(MC), hydroxypropyl methylcellulose (HPMC), ethyl cellulose (EC),hydroxyethyl cellulose (HEC), hydroxypropylcellulose (HPC),hydroxypropyl methylcellulose acetate, hydroxyethyl methylcellulose,hydroxyethylcellulose acetate, hydroxyethyl ethylcellulose andcombinations thereof.

Suitable emulsifiers include, but are not limited to, propylene glycolmonostearate (PGMS), sodium stearoyl lactylate (SSL), calcium stearoyllactylate (CSL), monoglycerides, diglycerides, monodiglycerides,polyglycerol esters, lactic acid esters, polysorbate, sucrose esters,diacetyl tartaric acid esters of mono-diglycerides (DATEM), citric acidesters of monoglycerides (CITREM) and combinations thereof. Additionalsuitable emulsifiers include DIMODAN, including DIMODAN™ B 727 andDIMODAN™ PV, GRINDSTED™ CITREM, GRINDSTED™ GA, GRINDSTED™ PS such asGRINDSTED™ PS 100, GRINDSTED™ PS 200, GRINDSTED™ PS 300, GRINDSTED™ PS400; RYLO™ (manufactured and distributed by DANISCO CULTOR), includingRYLO™ AC, RYLO™ CI, RYLO™ LA, RYLO™ MD, RYLO™ MG, RYLO™ PG, RYLO™ PR,RYLO™ SL, RYLO™ SO, RYLO™ TG; and combinations thereof.

Edible fibers include polysaccharides, oligosaccharides, lignin andassociated plant substances. Suitable edible fibers include, but are notlimited to, sugar beet fiber, apple fiber, pea fiber, wheat fiber, oatfiber, barley fiber, rye fiber, rice fiber, potato fiber, tomato fiber,other plant non-starch polysaccharide fiber, and combinations thereof.

Suitable flavoring agents include natural and synthetic flavors, “brownflavorings” (e.g., coffee, tea); dairy flavorings; fruit flavors;vanilla flavoring; essences; extracts; oleoresins; juice and drinkconcentrates; flavor building blocks (e.g., delta lactones, ketones);and the like; and combinations of such flavors. Examples of botanicflavors include, for example, tea (e.g., preferably black and greentea), aloe vera, guarana, ginseng, ginkgo, hawthorn, hibiscus, rosehips, chamomile, peppermint, fennel, ginger, licorice, lotus seed,schizandra, saw palmetto, sarsaparilla, safflower, St. John's Wort,curcuma, cardamom, nutmeg, cassia bark, buchu, cinnamon, jasmine, haw,chrysanthemum, water chestnut, sugar cane, lychee, bamboo shoots,vanilla, coffee, and the like.

Suitable sweeteners include, but are not limited to, alitame; dextrose;fructose; lactilol; polydextrose; xylitol; xylose; aspartame,saccharine, cyclamates, acesulfame K, L-aspartyl-L-phenylalanine loweralkyl ester sweeteners, L-aspartyl-D-alanine amides; L-aspartyl-D-serineamides; L-aspartyl-hydroxymethyl alkane amide sweeteners;L-aspartyl-1-hydroxyethylalkane amide sweeteners; and the like.

Suitable anti-oxidants include, but are not limited to, tocopherols(natural, synthetic); ascorbyl palmitate; gallates; butylatedhydroxyanisole (BHA); butylated hydroxytoluene (BHT); tert-butylhydroquinone (TBHQ); and the like.

Suitable coloring agents include, but are not limited to, FD&C dyes(e.g., yellow #5, blue #2, red #40), FD&C lakes; Riboflavin; β-carotene;natural coloring agents, including, for example, fruit, vegetable,and/or plant extracts such as grape, black currant, aronia, carrot,beetroot, red cabbage, and hibiscus.

Exemplary preservatives include sorbate, benzoate, and polyphosphatepreservatives.

Suitable emulsifiers include, but are not limited to, diglycerides;monoglycerides; acetic acid esters of mono- and diglycerides; diacetyltartaric acid esters of mono- and diglycerides; citric acid esters ofmono- and diglycerides; lactic acid esters of mono- and diglycerides;fatty acids; polyglycerol esters of fatty acids; propylene glycol estersof fatty acids; sorbitan monostearates; sorbitan tristearates; sodiumstearoyl lactylates; calcium stearoyl lactylates; and the like.

Suitable agents for pH adjustment include organic as well as inorganicedible acids. The acids can be present in their undissociated form or,alternatively, as their respective salts, for example, potassium orsodium hydrogen phosphate, potassium or sodium dihydrogen phosphatesalts. Exemplary acids are edible organic acids which include citricacid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconicacid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid andmixtures thereof.

Food Formulations

A subject formulation can be prepared in a variety of ways forconsumption by an individual, and, as indicated above, can include oneor more food components. Food formulations can be in a variety of forms,including powders; liquids; gels; and solid forms such as bars, tablets,capsules, candies, etc. Formulations of interest include foods forveterinary or human use, including food bars (e.g. cereal bars,breakfast bars, energy bars, nutritional bars); drinks; fortifieddrinks; carbonated beverages; drink supplements (e.g., powders to beadded to a drink); powders to be mixed with food; tablets; lozenges;candy; candy-like formulations, e.g., chewable gel formulations, e.g.,chewable gel candy in the shape of an animal; puddings; and the like.Suitable food formulations also include those described in U.S. Pat. No.7,067,150.

A food product can have final moisture content between about 0% andabout 100%, e.g., from about 0% to about 1%, from about 1% to about 5%,from about 5% to about 10%, from about 10% to about 30%, from about 30%to about 50%, from about 50% to about 80%, or from about 80% to about100%.

Packages

The present invention further provides a package comprising a subjectformulation. In some embodiments, a subject package comprises a singledosage form of a subject formulation. In other embodiments, a subjectpackage a subject package comprising multiple (e.g., 2, 3, 4, 5, 6, 7,8, 9, 10, or more) dosage forms of a subject formulation.

As one non-limiting example, a subject food product can be packaged insuch a way that multiple doses are contained in a single package,optionally where individual unit dosage forms are separated inindividual compartments in a single package. The dosage forms can be ina variety of forms, e.g., tablets or lozenges that are palatable (e.g.,flavored so as to be palatable, such as with fruit flavorings, sugars,and the like, as discussed above). Unit dosage forms include tablets,capsules, lozenges, candies, bars, a unit of powder (e.g., 1 tablespoonof a powder; a unit of a liquid, (e.g., a 1 tablespoon of a liquid),etc.

A subject package in some embodiments will further include instructionsfor use, including e.g., dosage amounts and dosage frequencies.Instructions are in some embodiments printed directly on the package. Inother embodiments, instructions are printed material provided as apackage insert. Instructions can also be provided in other media, e.g.,electronically in digital or analog form, e.g., on an audio cassette, anaudio tape, a compact disc, a digital versatile disk, and the like.

Exemplary Formulations

The following are exemplary formulations. As noted above, in addition tothe components specifically listed below, a subject formulation caninclude one or more additional active and/or inactive components,food-grade components, etc. In the exemplary formulations below, amountsare given as per unit dose.

Exemplary formulation 1 Component Amount EPA 700 mg DHA 250 mgα-tocopherol 500 mg γ-tocopherol 200 mg

Exemplary formulation 2 Component Amount EPA 700 mg DHA 250 mgα-tocopherol 1500 mg  γ-tocopherol 600 mg

In some embodiments, the amount of α-tocopherol in Exemplary formulation1 is increased to from 1000 mg to 3000 mg. In some embodiments, theamount of γ-tocopherol in Exemplary formulation 1 is increased to from500 mg to 1000 mg.

In some embodiments, the amount of α-tocopherol in Exemplary formulation2 is increased to from 1000 mg to 3000 mg. In some embodiments, theamount of γ-tocopherol in Exemplary formulation 2 is increased to from500 mg to 1000 mg.

In some embodiments, a subject formulation will include α-lipoic acidand/or carnitine.

Exemplary formulation 3 Component Amount EPA 700 mg DHA 250 mgα-tocopherol 500 mg γ-tocopherol 200 mg α-lipoic acid 100 mg

Exemplary formulation 4 Component Amount EPA 700 mg DHA 250 mgα-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoic acid 600 mg

Exemplary formulation 5 Component Amount EPA 700 mg DHA 165 mgα-tocopherol 500 mg γ-tocopherol 200 mg carnitine 200 mg

Exemplary formulation 6 Component Amount EPA 700 mg DHA 250 mgα-tocopherol 1500 mg  γ-tocopherol 600 mg carnitine 3000 mg 

In some embodiments, the amount of α-tocopherol in any one of Exemplaryformulations 3-6 is increased to from 1000 mg to 3000 mg. In someembodiments, the amount of γ-tocopherol in any one of Exemplaryformulations 3-6 is increased to from 500 mg to 1000 mg.

In some embodiments, a subject formulation will include an omega-6 fattyacid.

Exemplary formulation 7 Component Amount EPA 700 mg DHA 250 mg GLA  70mg α-tocopherol 500 mg γ-tocopherol 200 mg

Exemplary formulation 8 Component Amount EPA 700 mg DHA 250 mg GLA  70mg α-tocopherol 1500 mg  γ-tocopherol 600 mg

In some embodiments, a subject formulation will include an omega-9 fattyacid.

Exemplary formulation 9 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg

Exemplary formulation 10 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg

Exemplary formulation 11 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg

Exemplary formulation 12 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 600 mg

Exemplary formulation 13 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 200 mg

Exemplary formulation 14 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 600 mg Carnitine 3000 mg 

Exemplary formulation 15 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 200 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 1000 mg 

Exemplary formulation 16 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 200 mg Carnitine 3000 mg 

In some embodiments, a subject formulation includes one or more ofvitamin C, phosphocholine, zinc, and vitamin K. For example, exemplaryformulations include the following.

Exemplary formulation 17 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 1000 mg  Vitamin C 250 mg

Exemplary formulation 18 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 600 mg Carnitine 3000 mg  Vitamin C 500 mg

Exemplary formulation 19 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 200 mg Vitamin C 250 mg Phosphocholine  1 g

Exemplary formulation 20 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 600 mg Carnitine 3000 mg  Vitamin C 500 mg Phosphocholine 3 g

Exemplary formulation 21 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 200 mg Vitamin C 250 mg Phosphocholine  1 g Zinc 15 mg

Exemplary formulation 22 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 600 mg Carnitine 3000 mg  Vitamin C 500 mg Phosphocholine  3 g Zinc 30 mg

Exemplary formulation 23 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 200 mg Vitamin C 250 mg Phosphocholine  1 g Zinc 15 mg Vitamin K  5 μg

Exemplary formulation 24 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 250 mg α-tocopherol 1500 mg  γ-tocopherol 600 mg α-lipoicacid 600 mg Carnitine 3000 mg  Vitamin C 500 mg Phosphocholine  3 g Zinc30 mg Vitamin K  10 mg (K1 + K2)

Exemplary formulation 25 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 200 mg α-tocopherol 500 mg γ-tocopherol 200 mg α-lipoicacid 100 mg Carnitine 500 mg Vitamin C 100 mg Phosphocholine  1 g Zinc 5 mg Vitamin K1  3 mg Vitamin K2 350 μg

Exemplary formulation 26 Component Amount EPA 700 mg DHA 250 mg GLA  70mg Oleic acid 200 mg α-tocopherol 1500 mg  γ-tocopherol 500 mg α-lipoicacid 200 mg Carnitine 2000 mg  Vitamin C 500 mg Phosphocholine  3 g Zinc 15 mg Vitamin K1  10 mg Vitamin K2  1 mg

Exemplary formulation 27 Component Amount EPA 700 mg DHA 250 mg GLA 70mg Oleic acid 200 mg α-tocopherol 1500 mg γ-tocopherol 500 mg α-lipoicacid 200 mg Carnitine 2000 mg Vitamin C 500 mg Phosphocholine 3 g Zinc15 mg Vitamin K₁ 10 mg Vitamin K₂ 1 mg arginine 3000 mg Co-enzyme Q 200mg Selenium 50 μg Vitamin A 3000 IU Thiamin (B1) 7.5 mg Riboflavin (B2)7.5 mg Vitamin B6 200 mg Vitamin B12 1 mg Vitamin D 400 IU Calcium 100mg elemental magnesium 200 mg Biotin (B7) 50 μg Folic acid 400 μgPantothenic acid 10 mg tetrahydrobiopterin 50 mg Niacin 15 mg

A subject formulation can be prepared as a single dosage form, ordivided into two or more dosage forms. A subject formulation cancomprise one unit dose; two unit doses; three unit doses; or more thanthree unit doses. For example, any of exemplary formulations 1-27 can bedivided into two or more capsules, two or more tablets, two or morebars, two or more units of a powder (e.g., two or more grams of apowder, two or more tablespoons of a powder, etc.), two or more unitsliquid (e.g., two or more 1-ml units of a liquid, two or more 5-ml unitsof a liquid, etc.), two or more chewable gel units, or two or more unitsof another dosage form. Alternatively, a subject formulation can be aliquid formulation, where it can be formulated in a single dose (e.g.,1-15 ml) or formulated such that it is administered in two or moredoses, where each dose is 1-10 ml). For example, exemplary formulations1-27 provide the unit doses of the various components, which can beadministered to an individual in one, two, three, four, or more, doses,which doses can be in various dosage forms, e.g., tablets, capsules,liquids, powders, food products, etc.

Treatment Methods

The present invention provides methods of treating various disorders,involving administering to an individual in need thereof an effectiveamount of a subject formulation. A subject method is useful for treatingvarious disorders, including, but not limited to, apraxia, dyspraxia,autism, autism spectrum disorder, attention deficit/hyperactivitydisorder, dyslexia, depression, sensory integration dysfunction; immunesystem disorders such as celiac disease, sprue, gluten sensitivity, amalabsorption syndrome, asthma, food allergy, leaky gut syndrome, andeczema; cardiovascular disease; diabetes; and inflammatory conditionssuch as rheumatoid arthritis.

A subject formulation is in some embodiments administered to anindividual with apraxia, to treat the apraxia. In these embodiments, aneffective amount of a subject formulation is an amount that, whenadministered in one or more doses, is effective to provide for animprovement in one or more of an oral movement score, a simplephonemic/syllabic score, a complex phonemic/syllabic score, and aspontaneous length and complexity score, e.g., in the Kaufman PraxisTest. For example, an effective amount of a subject formulation is anamount that, when administered in one or more doses, is effective toprovide for an increase in percentile score of from about 5 to about 10percentile, from about 10 to about 20 percentile, from about 20 to about40 percentile, from about 40 to about 60 percentile, from about 60 toabout 70 percentile, or from about 70 to about 90 percentile, on one ormore of an oral movement score, a simple phonemic/syllabic score, acomplex phonemic/syllabic score, and a spontaneous length and complexityscore, e.g., in the Kaufman Praxis Test. Whether a subject formulationis effective in treating a disorder such as apraxia can be determinedusing well-established tests, such as the Kaufman Praxis Test.

A subject formulation is in some embodiments administered to anindividual with an immune system disorder, to treat the immune systemdisorder. In these embodiments, an effective amount of a subjectformulation is an amount that, when administered in one or more doses,is effective to provide for a reduction in one or more symptoms of animmune system disorder. For example, where the immune system disorder isan allergic disorder, an effective amount of a subject formulation is anamount that, when administered in one or more doses, is effective toprovide for a reduction in circulating factor that is an indicator of anallergic disorder of from about 10% to about 20%, from about 20% toabout 25%, from about 25% to about 30%, from about 30% to about 40%,from about 40% to about 50%, from about 50% to about 60%, from about 60%to about 70%, or from about 70% to about 80%, or more, of the level ofthe circulating factor present following exposure to anallergy-stimulating allergen. Circulating factors that are indicators ofan allergic disorder include, e.g., allergen-specific IgE; inflammatorybiomarkers; C-reactive protein; cytokines that are indicative of a Th2immune response; and the like. As another example, where the immunesystem disorder is an allergic disorder, an effective amount of asubject formulation is an amount that, when administered in one or moredoses, is effective to provide for one or more of: i) clinicalimprovement in one or more of sneezing, wheezing, runny nose, and othersymptoms of an allergic reaction; ii) decreased number of visits to amedical personnel for treatment for the allergic disorder; and iii)decreased use of medications used to treat the symptoms of an allergicdisorder. Whether a subject formulation is effective in treating animmune system disorder such as an allergic disorder can be determinedusing any well-established test, e.g., immunoassays for measuring IgElevels (e.g., allergen-specific IgE levels), and the like.

A subject formulation is administered, e.g., orally, to an individual inneed thereof in any frequency deemed appropriate to treat the conditionor disorder. For example, a subject formulation can be administeredthree times daily, twice daily, once daily, every other day, three timesper week, twice per week, once per week, or less often. In someembodiments, a subject formulation is administered daily. In otherembodiments, a subject formulation is administered every other day. Forexample, a unit dose of a subject formulation can be administered once,twice, or three times daily.

A subject formulation can be administered at any frequency, as discussedabove, over any period of time, as necessary to treat the condition ordisorder. Thus, a subject formulation can be administered over a periodof time of from about one week to about one month, from about one monthto about three months, from about three months to about 6 months, fromabout 6 months to about one year, from about one year to about threeyears, from about three years to about six years, from about six yearsto about 10 years, or more than 10 years.

The dosages of PUFA and vitamin E, as well as the dosages of additionalcomponents such as carnitine and α-lipoic acid, can vary according tovarious factors, including, e.g., the age of the individual, the weightof the individual, the genetic makeup of the individual, and theseverity of symptoms exhibited by the individual to whom a subjectformulation is administered. The following are general guidelines.

Where a subject formulation includes omega-3 fatty acids, the dosage ofthe omega-3 fatty acids ranges from about 100 mg/day to about 5000mg/day, e.g., from about 100 mg/day to about 200 mg/day, from about 200mg/day to about 300 mg/day, from about 300 mg/day to about 400 mg/day,from about 400 mg/day to about 500 mg/day, from about 500 mg/day toabout 600 mg/day, from about 600 mg/day to about 700 mg/day, from about700 mg/day to about 800 mg/day, from about 800 mg/day to about 900mg/day, from about 900 mg/day to about 1000 mg/day, from about 1000mg/day to about 1100 mg/day, from about 1100 mg/day to about 1200mg/day, from about 1200 mg/day to about 1300 mg/day, from about 1300mg/day to about 1400 mg/day, from about 1400 mg/day to about 1500mg/day, from about 1500 mg/day to about 2000 mg/day, from about 2000mg/day to about 3000 mg/day, from about 3000 mg/day to about 4000mg/day, or from about 4000 mg/day to about 5000 mg/day, where thedosages given are for individual omega-3 fatty acids or for totalomega-3 fatty acids (e.g., where more than one omega-3 fatty acid ispresent).

For example, in some embodiments, a subject formulation comprises theomega-3 fatty acids EPA and DHA. In some embodiments, the dosage for EPAwill range from about 500 mg/day to about 3000 mg/day, e.g., from about500 mg/day to about 600 mg/day, from about 600 mg/day to about 700mg/day, from about 700 mg/day to about 800 mg/day, from about 800 mg/dayto about 900 mg/day, or from about 900 mg/day to about 1000 mg/day; andthe dosage of DHA will range from about 100 mg/day to about 1000 mg/day,e.g., from about 100 mg/day to about 150 mg/day, from about 150 mg/dayto about 200 mg/day, from about 200 mg/day to about 250 mg/day, fromabout 250 mg/day to about 300 mg/day, from about 300 mg/day to about 350mg/day, from about 350 mg/day to about 400 mg/day, from about 400 mg/dayto about 500 mg/day, from about 500 mg/day to about 600 mg/day, fromabout 600 mg/day to about 700 mg/day, from about 700 mg/day to about 800mg/day, from about 800 mg/day to about 900 mg/day, or from about 900mg/day to about 1000 mg/day.

The dosage of α-tocopherol ranges from about 10 mg/kg/day to about 200mg/kg/day, e.g., from about 10 mg/kg/day to about 25 mg/kg/day, fromabout 25 mg/kg/day to about 50 mg/kg/day, from about 50 mg/kg/day toabout 75 mg/kg/day, from about 75 mg/kg/day to about 100 mg/kg/day, fromabout 100 mg/kg/day to about 125 mg/kg/day, from about 125 mg/kg/day toabout 150 mg/kg/day, from about 150 mg/kg/day to about 175 mg/kg/day, orfrom about 175 mg/kg/day to about 200 mg/kg/day; and the dosage ofγ-tocopherol ranges from about 100 mg/day to about 1000 mg/day, e.g.,from about 100 mg/day to about 200 mg/day, from about 200 mg/day toabout 300 mg/day, from about 300 mg/day to about 400 mg/day, from about400 mg/day to about 500 mg/day, from about 500 mg/day to about 600mg/day, from about 600 mg/day to about 700 mg/day, from about 700 mg/dayto about 800 mg/day, from about 800 mg/day to about 900 mg/day, or fromabout 900 mg/day to about 1000 mg/day.

The dosage of other forms of vitamin E (e.g., β-tocopherol,δ-tocopherol, α-tocotrienol, β-tocotrienol, δ-tocotrienol, andγ-tocotrienol), if present in a subject formulation, can range fromabout 5 mg/day to about 1000 mg/day, e.g., from about 5 mg/day to about10 mg/day, from about 10 mg/day to about 25 mg/day, from about 25 mg/dayto about 50 mg/day, from about 50 mg/day to about 75 mg/day, from about75 mg/day to about 100 mg/day, from about 100 mg/day to about 125mg/day, from about 125 mg/day to about 150 mg/day, from about 150 mg/dayto about 175 mg/day, from about 175 mg/day to about 200 mg/day, fromabout 200 mg/day to about 250 mg/day, from about 250 mg/day to about 500mg/day, from about 500 mg/day to about 750 mg/day, or from about 750mg/day to about 1000 mg/day, where the dosages given are for theindividual isoforms of vitamin E.

Where a subject formulation comprises one or more omega-6 fatty acids,the dosage of omega-6 fatty acid can range from about 50 mg/day to about1000 mg/day, e.g., from about 50 mg/day to about 75 mg/day, from about75 mg/day to about 100 mg/day, from about 100 mg/day to about 150mg/day, from about 150 mg/day to about 200 mg/day, from about 200 mg/dayto about 250 mg/day, from about 250 mg/day to about 300 mg/day, fromabout 300 mg/day to about 350 mg/day, from about 350 mg/day to about 400mg/day, from about 400 mg/day to about 450 mg/day, from about 450 mg/dayto about 500 mg/day, from about 500 mg/day to about 600 mg/day, fromabout 600 mg/day to about 700 mg/day, from about 700 mg/day to about 800mg/day, from about 800 mg/day to about 900 mg/day, or from about 900mg/day to about 1000 mg/day.

Where a subject formulation comprises one or more omega-9 fatty acids,the dosage of omega-9 fatty acid can range from about 50 mg/day to about500 mg/day, e.g., from about 50 mg/day to about 75 mg/day, from about 75mg/day to about 100 mg/day, from about 100 mg/day to about 150 mg/day,from about 150 mg/day to about 200 mg/day, from about 200 mg/day toabout 250 mg/day, from about 250 mg/day to about 300 mg/day, from about300 mg/day to about 350 mg/day, from about 350 mg/day to about 400mg/day, from about 400 mg/day to about 450 mg/day, or from about 450mg/day to about 500 mg/day.

Where a subject formulation comprises α-lipoic acid, the dosage ofα-lipoic acid can range from about 10 mg/kg/day to about 50 mg/kg/day,e.g., from about 10 mg/kg/day to about 20 mg/kg/day, from about 20mg/kg/day to about 25 mg/kg/day, from about 25 mg/kg/day to about 30mg/kg/day, from about 30 mg/kg/day to about 40 mg/kg/day, or from about40 mg/kg/day to about 50 mg/kg/day; or can range from about 50 mg/day toabout 1000 mg/day, e.g., from about 50 mg/day to about 75 mg/day, fromabout 75 mg/day to about 100 mg/day, from about 100 mg/day to about 150mg/day, from about 150 mg/day to about 200 mg/day, from about 200 mg/dayto about 250 mg/day, from about 250 mg/day to about 300 mg/day, fromabout 300 mg/day to about 350 mg/day, from about 350 mg/day to about 400mg/day, from about 400 mg/day to about 450 mg/day, from about 450 mg/dayto about 500 mg/day, from about 500 mg/day to about 600 mg/day, fromabout 600 mg/day to about 700 mg/day, from about 700 mg/day to about 800mg/day, from about 800 mg/day to about 900 mg/day, or from about 900mg/day to about 1000 mg/day.

Where a subject formulation comprises carnitine, the dosage of carnitinecan range from about 20 mg/kg/day to about 75 mg/kg/day, e.g., fromabout 20 mg/kg/day to about 25 mg/kg/day, from about 25 mg/kg/day toabout 30 mg/kg/day, from about 30 mg/kg/day to about 30 mg/kg/day, fromabout 40 mg/kg/day to about 50 mg/kg/day, from about 50 mg/kg/day toabout 60 mg/kg/day, or from about 60 mg/kg/day to about 75 mg/kg/day; orcan range from about 150 mg/day to about 3000 mg/day, e.g., from about150 mg/day to about 200 mg/day, from about 200 mg/day to about 250mg/day, from about 250 mg/day to about 300 mg/day, from about 300 mg/dayto about 350 mg/day, from about 350 mg/day to about 400 mg/day, fromabout 400 mg/day to about 450 mg/day, from about 450 mg/day to about 500mg/day, from about 500 mg/day to about 600 mg/day, from about 600 mg/dayto about 700 mg/day, from about 700 mg/day to about 800 mg/day, fromabout 800 mg/day to about 900 mg/day, from about 900 mg/day to about1000 mg/day, from about 1000 mg/day to about 1250 mg/day, from about1250 mg/day to about 1500 mg/day, from about 1500 mg/day to about 1750mg/day, from about 1750 mg/day to about 2000 mg/day, from about 2000mg/day to about 2250 mg/day, from about 2250 mg/day to about 2500mg/day, from about 2500 mg/day to about 2750 mg/day, or from about 2750mg/day to about 3000 mg/day.

Subjects Suitable for Treatment

Subjects suitable for treatment with a subject method includeindividuals who have been diagnosed with a disorder such as apraxia,dyspraxia, autism spectrum disorder, attention deficit/hyperactivitydisorder, dyslexia, or depression, sensory integration dysfunction; oran immune system disorder such as celiac disease, sprue, glutensensitivity, a malabsorption syndrome, asthma, food allergy, leaky gutsyndrome, or eczema; a cardiovascular disease; diabetes; or aninflammatory condition such as rheumatoid arthritis. Subjects suitablefor treatment with a subject method also include individuals who havebeen previously treated (with a treatment method other than a subjectmethod) for a disorder such as those listed above, but who have failedtreatment (e.g., failed to respond to the treatment), who have relapsed,or in whom the previous treatment method was contraindicated (e.g., dueto adverse reaction, etc.).

In some embodiments, the individual is a sub-adult human, e.g., aninfant (e.g., from about 1 month of age to about 12 months of age); atoddler (e.g., from about 12 months of age to about 3 years of age); apre-school age child (e.g., from about 3 years of age to about 5 yearsof age); a child from about 5 years of age to about 9 years of age; apre-teen from about 9 years of age to about 12 years of age; or ateenager (e.g., from about 13 years of age to about 19 years of age). Inother embodiments, the individual is an adult human, e.g., a human atleast 18-20 years old.

In some embodiments, the individual is a sub-adult human who has beendiagnosed with apraxia. In some embodiments, the individual is asub-adult human who has been diagnosed with autism spectrum disorder.

In some embodiments, the individual exhibits gluten sensitivity. In someembodiments, the individual has a human leukocyte antigen (HLA) allelethat is associated with gluten sensitivity. For example, in someembodiments, the individual has an HLA DQ1 allele. In some embodiments,the individual has elevated levels of anti-gliadin antibodies.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g. amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for. Unlessindicated otherwise, parts are parts by weight, molecular weight isweight average molecular weight, temperature is in degrees Celsius, andpressure is at or near atmospheric. Standard abbreviations may be used,e.g., bp, base pair(s); kb, kilobase(s); pl, picoliter(s); s or sec,second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); kb,kilobase(s); bp, base pair(s); nt, nucleotide(s); i.m.,intramuscular(ly); i.p., intraperitoneal(ly); s.c., subcutaneous(ly);and the like.

Example 1: Case Study Patient A

A boy with the diagnoses of asthma, gastro-esophageal reflux, multiplefood allergies, gluten-sensitivity (on a gluten/casein free diet sinceinfancy), and global apraxia/dyspraxia (severe expressive speechdisorder, and developmental coordination disorder—“clumsy childsyndrome”), hypotonia, right sided weakness, and sensory integrationdysfunction (poor propioception, abnormal vestibular sensation, and lackof appropriate pain sensation), chronic stearrhea, referred for earlyintervention evaluation and treatment at 17 months, responded slowly toconventional individual speech therapy, but learned quickly tocommunicate through sign language. At 23 months, he was non-verbal, atan expressive language level <9 months, frequently choked on his foodafter mouth stuffing, fell down 10-20 times and hour, often walking intowalls and tables, had very low tone in his upper extremities, decreaseddeep tendon reflexes, tactile defensiveness and no pain sensation in hisextremities. He was placed on omega 3/6/9 fatty acid therapy at 24months of age. This child remained nonverbal despite several months ofindividual speech therapy, however he demonstrated excellentprogress/improvement within days of initiation of PUFA supplementationin babbling, puckering, blowing bubbles, new word formation, andimproved coordination/less falling down. After 1 month of PUFAsupplementation, he experienced immediate regression within 48-72 hours(loss of speech ability and coordination) when fish oil supplements werestopped. He quickly regained these abilities when the PUFA supplementswere reinstated and surpassed them on higher doses. He has subsequentlydemonstrated resolution of all neurodevelopmental symptoms after theaddition of vitamin E (alpha and gamma).

Vitamin E (Vit E) supplementation was initiated after a literaturesearch revealed that its deficiency causes a constellation ofneurological symptoms that mirror those of apraxia of speech:articulation issues, poor coordination, low tone, and loss ofproprioception, vestibular and pain sensation. However this connectionhas not been previously made. Within 6 weeks of vitamin E+PUFAsupplements, this once severely apraxic/dyspraxic 36 month old childclinically presented as a normal 3 year old. Improvement in speechproduction/intelligibility and sentence length as well as improvedcoordination occurred rapidly. In addition a normalization of tone andsensory processing also occurred, including new processing of painsensation in his extremities, which was previously absent. Residualsymptoms of apraxia were apparent only on speech and OT assessments, andclinically with attempts at longer sentences and with tasks requiringcomplex motor planning.

The vit E dose was increased over several weeks and is now in the rangerecommended for neurological complications of vit E deficiency (100-200mg/kg/day alpha-d-tocopherol). Sokol et al. Gastroenterology 1993;104(6):1727-35. Plasma levels of alpha-d-tocopherol beforesupplementation fell within the range of normal (12.9 ng/L), and did notchange despite high dose supplementation. Currently at a total of 3000IU alpha and 600 IU gamma tocopherol a day, divided into 3 doses, he is90-100% intelligible and has experienced a complete resolution ofspeech, coordination and sensory abnormalities. All speech therapy andoccupational therapy has been phased out due to this rapid andremarkable improvement within 5 months of beginning therapy, at whichpoint it was felt by all evaluators to no longer be necessary.

Given that gut inflammation can cause malabsorption of key nutrientslike vitamin E, a link to food allergy and gluten-sensitivity/celiacdisease may exist. A metabolic work-up revealed a moderate/severecarnitine deficiency, a treatable condition that will impact fatty acidmetabolism that is commonly described in children with autistic spectrumdisorders. A secondary carnitine deficiency can occur in malabsorptionsyndromes like celiac disease. It is possible that gluten-sensitivityplaces children at increased risk for such neurological sequelae due tomalabsorption. HLA testing revealed the gluten sensitive HLA gene (DQB1*0503), shown to be associated with gluten ataxia and otherneurological complications of celiac disease. Carnitine supplementationwas initiated approximately 6 weeks after vit E supplementation hadbegun and significant improvement in tone, strength and coordination hasalready occurred. It is difficult to assess the additional contributionof carnitine supplementation to overall improvement. However givenfurther surges in speech and coordination with increased doses of vit E,the most significant contribution is felt to have come from vit E.

In addition to malabsorption, increased consumption of antioxidants dueto oxidative stress (Traber (2006) Br J Nutr 96 Suppl 1:S34-7; Bruno(2006) Pathophysiology 13(3):143-9; Brigelius-Flohe and Traber (1999)Faseb J 13(10):1145-55; Traber (1997) Eur J Clin Invest 27(10):822-4;Traber (1999) Vitamin E. In: Shils M E, Olson J A, al. E, editors.Modern Nutrition in Health and Disease. 9th ed. Baltimore: Williams andWilkens; p. 347-62) could also account for a potentially increasedutilization of vit E and lack of change in plasma alpha tocopherollevels despite high-dose supplementation in this child. Abnormalities inalpha tocopherol transport into the brain and nervous system is anothermechanism that could account for the neurological complications despiteapparently adequate plasma levels. Genetic abnormalities of the alphatocopherol transport protein have been described and are associated withneurological complications. Sokol R. Vitamin E deficiency andneurological disorders. In: Pacher L, Fuchs J, editors. Vitamin E inHealth and Disease. New York: Marel Dekker; 1993. p. 815-49. Although atocopherol transport protein has not yet been identified in the brain,it is assumed to exist. Regardless of the underlying mechanism, simplesupplementation has led to a clinical cure in this child. However theunderlying condition remains, as neurological symptoms rapidly reoccureven with a slight decrease in dosage of vit E supplementation.

The discovery of a moderate to severe carnitine deficiency may also bethe result of a mild mitochondrial disorder. Subsequently additionalantioxidants often part of a mitochondrial cocktail have been added toPatient A's daily regimen, including co-enzyme Q (100 mg/day), vitamin C(250 mg/day) and alpha-lipoic acid (300 mg/day) resulting in increasedclarity of speech and improved energy level/stamina within 1 week ofsupplementation. Polyenylphosphatidylcholine (3 grams/day) is a moleculefound in lecithin or egg believed to increase choline levels in theblood and brain and supports acetylcholine synthesis for proper neuronaland cell function and provides gastric mucosal protection. Decreasedinflammation has been reported with its use, (Demirbilik et al,Intensive Care Med 2004, 30:1974-8), which may be of benefit indisorders involving increased oxidative stress. Patient A experienced adecreased in stearrhea within a week of supplementation, suggestingimprovement in fat absorption. In addition Vitamin K (to prevent adverseeffect of vitamin K antagonism by high dose vitamin E), a completemulti-vitamin combination (with selenium, zinc and high B-complex andadditional fat soluble vitamins) was also added to the daily regimen.Although the most dramatic effects on symptoms of apraxia occurred withthe use of omega 3+alpha and gamma tocopherol, the combination ofcarnitine, additional antioxidants and nutritional supplements let toglobal improvement in speech, coordination and sensory processing forPatient A that can not be explained by the intervention of speech andoccupational therapy alone. Patient A has achieved a clinical cure ofglobal apraxia with the above interventions. However during a recentillness of acute gastroenteritis that involved 2 days of vomiting whichprohibited ingestion of the multiple nutritional supplements of thisapraxia regimen, Patient A began to lose speech clarity/intelligibilityand again began to demonstrate poor coordination with more frequentfalls. The regression off supplements quickly resolved within 2-3 days,once the vomiting ceased and supplements were reintroduced. Dramaticneurological regressions have also been noted to occur with spoiled(oxidized/rancid) omega 3, benign viral illness stressors and conditionsthat trigger increased inflammation, including exposures to known foodallergens in patient A (milk, gluten, yeast, mold, fungus). Specificfood allergies have been confirmed through skin prick and patch testing,RAST testing and elimination diets, working closely with a pediatricallergist. Subsequent improvement in speech, coordination and behaviorwas noted with use of oral prednisone or dexamethasone for treatment ofallergic reactions and asthma exacerbations triggered by food allergy.Further neurologic benefits were noted when the antifungal nystatin wasused to treat oral candidiasis (from use of inhaled steroids as asthmatherapy). Allergy to bakers/brewers yeast, mold and aspergillus wassubsequently identified. Benefits from antifungals in this patient'scase could be the result of decreased allergen load/exposure in a childwith a yeast allergy.

Interventions: Speech Therapy and Occupational Therapy

At 21 months, 1:1 individual speech therapy (ST) was initiated at 60minutes once a week and an additional 60 minutes once a week was spentworking with an early intervention specialist who came to the home. STwas increased to 3 times a week between 27-36 months, and occupationaltherapy (OT) (1 hour) was started at twice a week for global dyspraxia,hypotonia and right-sided weakness. Steady improvement was made,especially after the addition of PUFA supplementation. However the childremained moderately apraxic/dyspraxic.

The Kaufman Speech Praxis Test (Table 1) was performed by a qualifiedspeech and language pathologist at 24 month with scores in the ≤3^(rd)percentile in all areas tested, demonstrating severe apraxia of speech.It was repeated 1 year later, approximately 4 weeks after vit E therapywas initiated. Overall intelligibility based on a 200 utterancecollection to a familiar listener was calculated at 60% (previously<25%), <50% by a stranger in known contexts (previously >10%), <40% bystranger in unknown contexts (previously <10%). Mean length of utterancewas calculated at 3.2 words. Although he still maintained a diagnosis ofmoderate apraxia that became more prominent with longer and more complexattempts at conversation, he made rapid, dramatic improved within 4weeks of initiation of vit E supplementation. At the 36-month transitionfrom regional center, this child still qualified for speech therapy (1hour group) through the school district twice a week. Private 1:1 ST (1hour) was planned to continue twice a week. Remarkable globalimprovement continued on vit E therapy as doses were increased. By laterthat same year, the apraxia diagnosis was no longer applicable. He was90-100% intelligible even to strangers, and it was felt that ST and OTwere no longer necessary, and school district-based services werecancelled. Repeat Kaufman testing reveals the highest attainable scoresin all fields. Sentence structure on was calculated at the 99^(th)percentile, and speech therapy was discontinued.

TABLE 1 Improvements in Kaufman Praxis Test: Standard score (Percentile)Pre Vitamin E 4 wk Tx Vit E 16 wk Tx Vit E Kaufman Categories (Age: 24mos) (Age: 35 mos) (Age: 38 mos) Oral Movement 72 (3^(rd))  97 (28^(th))110 (40^(th))* Simple Phonemic/ 12 (<3^(rd)) 106 (42^(nd))* Highestscore Syllabic already achieved Complex 56 (<3^(rd))  83 (16^(th)) 117(97^(th))* Phonemic/Syllabic Spontaneous 69 (3^(rd))  80 (9^(th)) 111(58^(th))* Length & Complexity

Example 2: Impact of Vitamin E and Omega 3 Supplementation in Childrenwith Verbal Apraxia

Verbal apraxia (VA) is a neurologically-based motor planning disorder ofunknown etiology common in autism spectrum disorders (ASD) thatanecdotally responds to omega 3 polyunsaturated fatty acid (PUFA)supplementation. Vitamin (vit) E deficiency causes symptoms that overlapthose of VA. PUFAs in the cell membrane are vulnerable to lipidperoxidation and early destruction if vit E is not readily available,potentially leading to neurological sequelae. Inflammation of thegastrointestinal tract and gluten sensitivity may contribute tomalabsorption of nutrients such as vit E and carnitine, contributing tofatty acid metabolism dysfunction and neurological abnormalities.

Objective:

Determine efficacy of vit E and PUFA supplementation in children withVA.

Design/Methods:

50 children diagnosed with VA were treated with vit E+PUFA. 10 of thesechildren were known to have ASD. A celiac panel, fat soluble vitamins,and carnitine level were obtained in patients having blood analyzed.

Results:

Age ranged from 2-13 years, (majority <5 yrs), and 38/50 were boys. Ahistory of gastrointestinal symptoms, sensory integration dysfunction,low muscle tone and coordination difficulties (dyspraxia) was commonlyreported. Forty-eight families (96%) anecdotally reported dramaticimprovements in a number of areas including speech, imitation,coordination, eye contact, behavior, sensory issues and the developmentof pain sensation. Two children experienced new tearful or aggressivebehavior within 3 days of initiating vit E (400 IU/d) without apparentbenefits in speech, and therapy was withdraw within a week. No otheradverse effects were reported. Plasma alpha tocopherol levels varied inchildren tested (low in 2, high in 4, and normal in 4); howeverpre-treatment levels did not reflect clinical response. Low plasmacarnitine was identified in 13/14 (93%) children. Antigliadin IgGantibodies were high in 9/11 (82%) children tested gluten-sensitivityHLA alleles were identified in 7 out of 7 boys screened. Two childrenreported vit D deficiency and early signs of rickets, and zincdeficiency was identified in 2 children. Abnormal fecal fat stoolstudies done in 4 children identified fat malabsorption in all 4children.

Conclusions:

Described here is a new disease paradigm of abnormal vit E and fattyacid metabolism causing neurological dysfunction in VA that responds toa safe nutritional intervention. The association of carnitine deficiencyand gluten sensitivity with VA is a novel observation, suggesting thatthese children deserve a more comprehensive metabolic work-up than whatis current standard practice.

Example 3: Characterization of a Neurodevelopmental Condition thatResponds to Omega-3 and Vitamin E Supplementation

The symptoms and metabolic anomalies of a cohort of children with VAthat may respond to nutritional interventions were characterized. 183children with VA (age range 2-15; median age 4 yrs, >20% ASD), weretreated with vit E+PUFA. A celiac panel, fat soluble vitamins, andcarnitine level was obtained in patients having blood analyzed. A commonclinical phenotype of male predominance (80%), GI symptoms (diarrhea,constipation, abdominal pain, vomiting, gastroesophageal reflux), foodallergy (skin-test/patch test/RAST test positive), sensory issues, lowmuscle tone and coordination difficulties emerged. 177 families (97%)anecdotally reported dramatic improvements in a number of areasincluding speech, imitation, coordination, eye contact, behavior,sensory issues and the development of pain sensation. Doses of vit Eused range from 400 IU-3000 IU a day, with the majority of familiesusing 800 IU a day divided into 2 doses, and an additional gammatocopherol supplement (200-400 mg/day). No adverse events have occurred.A variety of blood testing was performed and reported in 23 children,with a summary of abnormal laboratory results described below.

1. Plasma Alpha-d-Tocopherol Levels:

Broad range of levels reported in 12 children, both low (n=3), high(n=4) and normal (n=5) prior to supplementation. Pre-supplementationlevels do not appear to reflect clinical response to vit E.

2. Plasma Carnitine Levels:

Low plasma carnitine (total & free) was reported in 19/24 (79%)children. Total carnitine levels were often 50-70% below the lower limitof normal, suggesting a moderate to severe deficiency. Although acarnitine deficiency is not uncommon in children with ASD, this is anovel observation in apraxia.

3. Antigliadin IgG Antibodies:

High in 15/19 (79%) children tested (8 had HLA testing done, and 100%revealed the presence of a DQ1 “gluten-sensitivity” gene, or celiac HLADQ2).

4. Fat Soluble Vitamins:

3/4 reports of vitamin D deficiency, and early signs of rickets in 2children. One child had signs of rickets identified on wrist films.

5. Plasma Selenium:

One out of 2 boys tested had a significant selenium deficiency.

6. HLA Testing:

Performed on 8 boys, 100% of whom carried the “gluten-sensitivity” HLA(5 with DQ1 gene, known to be associated with neurological complicationsof gluten-sensitivity, and 3 with DQ2 associated with classic celiacdisease). The frequency of this HLA genotype is high in our cohort,given its prevalence of about 30% in Caucasians. Antigliadin IgGantibodies were elevated in 7 of these boys on a gluten-containing diet.The seventh child, negative for antigliadin antibodies, had beengluten-free since infancy due to severe abdominal symptoms and a familyhistory of gluten-intolerance. Intestinal biopsies done on 4 boys werenegative for “classic” celiac disease, although one demonstrated villousatrophy.

7. Fat Malabsorption Syndrome:

Identified on 7/9 boys on qualitative fecal fat studies.

8. Erythrocyte Glutamine/Glutamate Ratio:

Novel biomarker of oxidative stress (Morris et al Blood 2008), was verylow in the one apraxic patient measured (prior to vit E therapy),suggesting significant oxidative stress with a ratio lower than the mostill patients with sickle cell disease (480 μM/241 μM); ratio=2(normal=5.5±1.3).

9. Leukocyte Redox Studies:

Low GSH and altered metabolism in immune cells in autism vs., normalcontrols has been documented (Jung et al, AJBB 2008), as noted by theSulfur Amino Acid data below. In comparison, GSH is remarkably high inthe apraxic and dyspraxic patients (n=2) after 1 year of PUFA/Vit E.Given the critical role of glutathione in maintaining redox balance andhandling oxidative stress, a therapy that increases glutathionebioavailability is desirable.

Control Autism (n = 11) (n = 30) Dypraxic Pt Apraxic Pt Total Met (Met +Met-So) 0.9 ± 0.3 1.3 ± 0.8 0.13841 0.12523 SAM 0.043 ± 0.02  0.028 ±0.02  0.04670 0.03970 SAH 0.0096 ± 0.009  0.012 ± 0.008 0.01280 0.00700SAM/SAH ratio 6.5 ± 4.6 3.3 ± 2.8 3.64844 5.67143 Total Cys (Cys + (2 *Cys2)) 0.27 ± 0.12 0.16 ± 0.11 0.49 0.56 Total GSH (GSH + (2 * GSSG))2.14 1.59 3.97 3.14 Glutamine  0.4 ± 0.17 0.34 ± 0.24 1.11 1.05Glutamate  0.6 ± 0.26 0.75 ± 0.92 1.17 2.34 Gln/Glu ratio 0.67 ± 0.2 0.49 ± 0.56 0.94 0.45

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

What is claimed is:
 1. A method of treating apraxia and/or autism spectrum disorder, the method comprising orally administering to an individual in need thereof an effective amount of a formulation comprising: a) eicosapentaenoic acid (EPA); b) docosahexaenoic acid (DHA); c) α-tocopherol; and d) γ-tocopherol, wherein the ratio of EPA to DHA is in a range of from about 1.5:1 to about 5:1, wherein the EPA is present in an amount of from 500 mg to 3000 mg per unit dose, wherein the DHA is present in an amount of from 100 mg to 400 mg per unit dose, wherein the α-tocopherol is present in an amount of from about 300 mg to about 3000 mg per unit dose, and wherein the γ-tocopherol is present in an amount of from about 100 mg to about 1000 mg per unit dose.
 2. The method of claim 1, wherein the formulation is administered three times daily.
 3. The method of claim 1, wherein the formulation is administered twice daily.
 4. The method of claim 1, wherein the formulation is administered once daily.
 5. The method of claim 1, wherein said administration is effective to increase the percentile score of at least one of oral movement score, simple phonemic/syllabic score, complex phonemic/syllabic score, and spontaneous length and complexity score.
 6. The method of claim 1, wherein the formulation further comprises α-lipoic acid in an amount of from about 50 mg to about 600 mg per unit dose.
 7. The method of claim 1, wherein the formulation further comprises carnitine in an amount of from about 200 mg to about 3000 mg per unit dose.
 8. The method of claim 1, wherein the formulation further comprises an omega-6 fatty acid.
 9. The method of claim 8, wherein the omega-6 fatty acid is γ-linolenic acid.
 10. The method of claim 1, wherein the formulation further comprises an omega-9 fatty acid.
 11. The method of claim 10, wherein the omega-9 fatty acid is oleic acid.
 12. The method of claim 1, wherein the formulation further comprises vitamin C in an amount of from about 200 mg to about 500 mg.
 13. The method of claim 1, wherein the formulation further comprises vitamin K.
 14. The method of claim 1, wherein the formulation further comprises phosphocholine.
 15. The method of claim 1, wherein the formulation further comprises zinc.
 16. The method of claim 1, wherein the formulation further comprises one or more additional components selected from coenzyme Q, selenium, vitamin A, vitamin B₁, riboflavin, vitamin B₆, vitamin B₁₂, vitamin D, arginine, calcium, magnesium, vitamin B₇, vitamin B₉, vitamin B₅, tetrahydrobiopterin, and vitamin B₃.
 17. The method of claim 1, wherein the formulation further comprises a pancreatic enzyme.
 18. The method of claim 1, wherein the formulation further comprises a leukotriene inhibitor.
 19. The method of claim 1, wherein the formulation is in a dosage form selected from a tablet, a capsule, a powder, a gel, and a liquid.
 20. The method of claim 1, wherein the formulation further comprises one or more food-grade components fit for human consumption.
 21. The method of claim 9, wherein the γ-linolenic acid is present in an amount of from about 50 mg to about 75 mg.
 22. The method of claim 1, wherein the formulation further comprises: vitamin K1 in an amount of from 100 μg to 10 mg; and vitamin K2 in an amount of from 100 μg to 2 mg.
 23. The method of claim 1, wherein the formulation further comprises vitamin D3.
 24. The method of claim 23, wherein the vitamin D3 is present in an amount of from 200 IU to 800 IU.
 25. The method of claim 1, wherein the formulation further comprises an anti-oxidant.
 26. The method of claim 1, wherein the formulation further comprises an anti-inflammatory agent.
 27. The method of claim 1, wherein the formulation further comprises an amino acid. 